Anti-IL-5 antibodies

Abstract

Disclosed herein are fully human antibody molecules that immunospecifically bind to human IL-5. The antibody molecules can bind to human IL-5 with an equilibrium affinity constant (KD) of at least about 40 pM as determined by surface plasmon resonance.

Description

CROSS REFERENCE TO RELATED APPLICATIONS

[0001] This application is a continuation of U.S. application Ser. No. 15 / 848,090, filed Dec. 20, 2017 (now U.S. Pat. No. 11,111,292), which claims priority to U.S. Provisional Application No. 62 / 438,502, filed Dec. 23, 2016, the disclosures of each of which are hereby incorporated by reference in their entirety.SEQUENCE LISTING

[0002] The instant application contains a sequence listing which has been submitted electronically in ASCII format and is hereby incorporated by reference in its entirety. Said ASCII copy, created on Jul. 26, 2021, is named 102085.001560_sl.txt and is 91 KB in size.TECHNICAL FIELD

[0003] Disclosed herein are novel antibody molecules that immunospecifically bind IL-5, and uses of the disclosed antibodies.BACKGROUND

[0004] Interleukin-5 (IL-5) is a T-helper 2 (Th2) cytokine which causes the proliferation and differentiation of both B cells and eosinophils. In B cells, IL-5 also increases immunoglobulin secretion. IL-5 is a key modulator of eosinophils, where it also regulates maturation, migration to tissues, survival, and the prevention of apoptosis.

[0005] Through two separate motifs, IL-5 binds to its specific receptor (IL5-Rα) and a signaling receptor, a common β-chain (βc) shared between Interleukin-3 (IL-3) and granulocyte-macrophage colony stimulating factor (GMCSF). The affinity of IL-5 for IL5-Rα has been reported to be in the mid-low nM range (0.2-100 nM); this shifts into the mid-pM range (˜100 pM) in the presence of βc. IL5-Rα binds to IL-5 specifically, which then recruits βc to IL-5R.

[0006] The therapeutic potential of targeting interleukin-5 (IL-5) has been demonstrated by extensive validation in the literature and recent positive Phase III clinical data for both reslizumab and mepolizumab.SUMMARY

[0007] Disclosed herein are human antibody molecules that immunospecifically bind to human IL-5 with an equilibrium affinity constant (KD) of at least about 40 pM as determined by surface plasmon resonance.

[0008] Also provided are antibody molecules comprising a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO: 6, a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO: 8, a light chain CDR1 comprising the amino acid sequence of SEQ ID NOs: 5, 21, 24, 27, 30, 33, 36, 39, or 66, a light chain CDR2 comprising the amino acid sequence of SEQ ID NOs: 7, 42, or 45, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NOs: 15, 48, 51, 54, 57, 60, or 63.

[0009] The disclosed antibody molecules can comprise a heavy chain variable region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 16 and a light chain variable region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 17, 22, 25, 28, 31, 34, 37, 40, 43, 46, 49, 52, 55, 58, 61, 64, or 67, wherein the variability (i.e. the at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identity) occurs outside of the CDR sequence.

[0010] The disclosed antibody molecules can comprise a heavy chain comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NOs: 18 or 20 and a light chain comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 19, 23, 26, 29, 32, 35, 38, 41, 44, 47, 50, 53, 56, 59, 62, 65, or 68, wherein the variability (i.e. the at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identity) occurs outside of the CDR sequence.

[0011] Nucleic acid molecules encoding the disclosed antibody molecules, vectors comprising the nucleic acid molecules, and cells transformed to express the disclosed antibody molecules are also provided.

[0012] Also disclosed are pharmaceutical compositions comprising any of the antibody molecules disclosed herein.

[0013] Also provided are methods of treating a subject having eosinophilic asthma, hypereosinophilic syndrome, nasal polyposis with eosinophilic involvement, eosinophilic granulomatosis with polyangiitis, atopic dermatitis or eosinophilic esophagitis comprising administering to the subject a therapeutically effective amount of any of the herein disclosed antibody molecules, or pharmaceutical compositions comprising the same, to treat the eosinophilic asthma, hypereosinophilic syndrome, nasal polyposis with eosinophilic involvement, eosinophilic granulomatosis with polyangiitis, atopic dermatitis or eosinophilic esophagitis.

[0014] Use of an effective amount of any of the herein disclosed antibody molecules, or pharmaceutical compositions comprising the same, in the treatment of eosinophilic asthma, hypereosinophilic syndrome, nasal polyposis with eosinophilic involvement, eosinophilic granulomatosis with polyangiitis, atopic dermatitis, or eosinophilic esophagitis are also provided.

[0015] Further provided is the use of any of the herein disclosed antibody molecules, or pharmaceutical compositions comprising the same, in the manufacture of a medicament for the treatment of eosinophilic asthma, hypereosinophilic syndrome, nasal polyposis with eosinophilic involvement, eosinophilic granulomatosis with polyangiitis, atopic dermatitis or eosinophilic esophagitis.BRIEF DESCRIPTION OF THE DRAWINGS

[0016] The summary, as well as the following detailed description, is further understood when read in conjunction with the appended drawings. For the purpose of illustrating the disclosed antibody molecules, methods, and uses, there are shown in the drawings exemplary embodiments of the antibody molecules, methods, and uses; however, the antibody molecules, methods, and uses are not limited to the specific embodiments disclosed. In the drawings:

[0017] FIG. 1 illustrates a TF-1.6G4 assay showing that antibody variant 3A5.001 (in IgG4 format) retained an equivalent potency to the original antibody 3A5 (in IgG1 format).

[0018] FIG. 2 illustrates a TF-1.6G4 assay showing that antibody variant 3A5.040 retained an equivalent potency to the parent antibody 3A5.001.

[0019] FIG. 3 is a graphical matrix showing the position and identity of the different single amino acid substitutions that were generated on the 3A5.040 VH CDRs, aligned to the original 3A5.040 VH sequence (top sequence). The boxes contain the residues which were designated CDR residues according to the AbM nomenclature. In addition to the CDR residues, variants were made to Kabat residue numbers 93 and 94 (adjacent to HC CDR3) as described in the Example “Functional testing and characterization of antibodies.” The various sequences recited in this figure are provided in SEQ ID NO:73.

[0020] FIG. 4 is graphical matrix showing the position and identity of the different single amino acid substitutions that were generated to the 3A5.040 VL CDRs, aligned to the original 3A5.040 VL sequence (top sequence). The boxes contain the residues which were designated CDR residues according to the AbM nomenclature. The various sequences recited in this figure are provided in SEQ ID NO:74.

[0021] FIG. 5 illustrates exemplary light chain CDR consensus sequences aligned to the 3A5.040 VL sequence (top sequence). 3A5.040 VL CDR single amino acid substitution variants which showed potential improvements according to the inclusion criteria described in the Examples of (the ratio of variant kd to 3A5.040 kd≥1.5) and (the ratio of variant expression level to 3A5.040 expression level ≥0.5) were included in the consensus sequences. CDR definitions and numbering are according to AbM and Kabat nomenclature, respectively. The boxes identify the positions of CDR residues. The various sequences recited in this figure are provided in SEQ ID NO:75.

[0022] FIG. 6 illustrates a multi-concentration BIACORE™ kinetic analysis of 3A5.046 antibody binding to recombinant human IL-5 at 2.5, 1.25, 0.625, 0.313 and 0.156 μg / mL. Sensorgrams show decreasing concentrations of IL-5 in order from 2.5 μg / mL IL-5 in the top trace to 0.156 μg / mL IL-5 in the bottom trace.

[0023] FIG. 7A and FIG. 7B illustrate the proliferation of TF-1.6G4 cells in response to IL-5, and the potency of inhibition of IL-5 driven proliferation by 3A5.046. 3A5.046 was a more potent inhibitor of human IL-5 (FIG. 7A) and cynomolgus monkey IL-5 (FIG. 7B) than mepolizumab.

[0024] FIG. 8A and FIG. 8B illustrate an exemplary ELISA developed using the 3A5 antibody and a control capture antibody (R&D SYSTEMS®). The ELISA was able to detect recombinant IL-5 (FIG. 8A) and IL-5 produced from CD3 / CD28 / IL-33 activated primary human T-cells from 3 donors (FIG. 8B). FIG. 8B top panel: Donor 1, FIG. 8B middle panel: Donor 3; FIG. 8B bottom panel: Donor 4.

[0025] FIG. 9 illustrates the results of an experiment in which CD34+ cord blood cells differentiated into phenotypically mature eosinophils using IL-5 and other cytokines as described in the Examples. Antibody 3A5.046 was more potent than mepolizumab at inhibiting IL-5 induced eosinophil differentiation.

[0026] FIG. 10A, FIG. 10B, FIG. 10C, FIG. 10D, and FIG. 10E illustrate the results of the BIACORE™ crossreactivity analysis of antibody 3A5.046 binding to human IL-5 (FIG. 10A), cynomolgus monkey IL-5 (FIG. 10B), mouse IL-5 (FIG. 0C), rat IL-5 (FIG. 10D), and guinea pig IL-5 (FIG. 10E). Double-referenced sensorgrams are shown for binding to cytokines at 1 μg / ml or 10 μg / ml.

[0027] FIG. 11A, FIG. 11B, FIG. 11C, and FIG. 11D illustrate the results of a BIACORE™ specificity analysis of antibody 3A5.046. Double-referenced sensorgrams are shown for binding to cytokines at 10 μg / ml (FIG. 11A and FIG. 11B) or 1 μg / ml (FIG. 11C and FIG. 11D).

[0028] FIG. 12 illustrates the results of Ascaris suum (A. suum) challenge in a cynomolgus monkey model of airway eosinophilia. At day 2 there was a substantial difference in lung (BALF) eosinophil numbers when comparing 3A5.046-treated and vehicle (placebo)-treated animals (p<0.01; Mann-Whitney test).

[0029] FIG. 13A and FIG. 13B illustrate the blood eosinophil response over 10 days following an A. suum challenge in cynomolgus monkeys which were pre-treated with 3A5.046 or a isotype-matched control antibody (“placebo”) one week before A. suum challenge (FIG. 13A). FIG. 13B illustrates further details of the blood eosinophil counts for animals treated with 3A5.046 antibody, up to 45 days post-challenge (52 days post-dose). Eosinophil counts remained below baseline for at least 45 days post-challenge following a single dose of 3A5.046 one week before challenge.

[0030] FIG. 14 illustrates BALF eosinophil levels in a human IL-5 knock-in (KI) rat model in response to a challenge with Alternaria alternata. This model may be used to test the potency of an anti-IL-5 antibody to modulate the numbers of BALF eosinophils following Alternaria challenge.DETAILED DESCRIPTION OF ILLUSTRATIVE EMBODIMENTS

[0031] The disclosed antibody molecules, methods, and uses may be understood more readily by reference to the following detailed description taken in connection with the accompanying figures, which form a part of this disclosure. It is to be understood that the disclosed antibody molecules, methods, and uses are not limited to the specific antibody molecules, methods, and uses described and / or shown herein, and that the terminology used herein is for the purpose of describing particular embodiments by way of example only and is not intended to be limiting of the claimed antibody molecules, methods, and uses.

[0032] Unless specifically stated otherwise, any description as to a possible mechanism or mode of action or reason for improvement is meant to be illustrative only, and the disclosed antibody molecules, methods, and uses are not to be constrained by the correctness or incorrectness of any such suggested mechanism or mode of action or reason for improvement.

[0033] Throughout this text, the descriptions refer to antibody molecules and methods of using said antibody molecules. Where the disclosure describes or claims a feature or embodiment associated with an antibody molecule, such a feature or embodiment is equally applicable to the methods of using said antibody molecule. Likewise, where the disclosure describes or claims a feature or embodiment associated with a method of using an antibody molecule, such a feature or embodiment is equally applicable to the antibody molecule.

[0034] Where a range of numerical values is recited or established herein, the range includes the endpoints thereof and all the individual integers and fractions within the range, and also includes each of the narrower ranges therein formed by all the various possible combinations of those endpoints and internal integers and fractions to form subgroups of the larger group of values within the stated range to the same extent as if each of those narrower ranges was explicitly recited. Where a range of numerical values is stated herein as being greater than a stated value, the range is nevertheless finite and is bounded on its upper end by a value that is operable within the context of the invention as described herein. Where a range of numerical values is stated herein as being less than a stated value, the range is nevertheless bounded on its lower end by a non-zero value. It is not intended that the scope of the invention be limited to the specific values recited when defining a range. All ranges are inclusive and combinable.

[0035] Reference to a particular numerical value includes at least that particular value, unless the context clearly dictates otherwise.

[0036] When values are expressed as approximations, by use of the antecedent “about,” it will be understood that the particular value forms another embodiment. The term “about” when used in reference to numerical ranges, cutoffs, or specific values is used to indicate that the recited values may vary by up to as much as 10% from the listed value. Thus, the term “about” is used to encompass variations of ±10% or less, variations of ±5% or less, variations of ±1% or less, variations of ±0.5% or less, or variations of ±0.1% or less from the specified value.

[0037] It is to be appreciated that certain features of the disclosed antibody molecules, methods, and uses which are, for clarity, described herein in the context of separate embodiments, may also be provided in combination in a single embodiment. Conversely, various features of the disclosed antibody molecules, methods, and uses that are, for brevity, described in the context of a single embodiment, may also be provided separately or in any subcombination.

[0038] As used herein, the singular forms “a,”“an,” and “the” include the plural.

[0039] Various terms relating to aspects of the description are used throughout the specification and claims. Such terms are to be given their ordinary meaning in the art unless otherwise indicated. Other specifically defined terms are to be construed in a manner consistent with the definitions provided herein.

[0040] The term “comprising” is intended to include, but is not necessarily limited to, examples encompassed by the terms “consisting essentially of” and “consisting of”; similarly, the term “consisting essentially of” is intended to include, but is not necessarily limited to, examples encompassed by the term “consisting of.”

[0041] The term “antibody molecule” is meant in a broad sense and includes full length immunoglobulin molecules and antigen binding fragments thereof.

[0042] Immunoglobulins can be assigned to five major classes, namely IgA, IgD, IgE, IgG, and IgM, depending on the heavy chain constant domain amino acid sequence. IgA and IgG are further sub-classified as the isotypes IgAQ1, IgA2, IgG1, IgG2, IgG3, and IgG4. Antibody light chains of any vertebrate species can be assigned to one of two clearly distinct types, namely kappa (x) and lambda (k), based on the amino acid sequences of their constant domains.

[0043] “Antigen binding fragment” refers to a portion of an immunoglobulin molecule that retains the antigen binding properties of the parental full length antibody (i.e. “antigen binding fragment thereof”). Exemplary antigen binding fragments can have: heavy chain complementarity determining regions (HCDR) 1, 2, and / or 3; light chain complementarity determining regions (LCDR) 1, 2, and / or 3; a heavy chain variable region (VH); a light chain variable region (VL); and combinations thereof. Antigen binding fragments include: a Fab fragment, a monovalent fragment consisting of the VL, VH, constant light (CL), and constant heavy 1 (CH1) domains; a F(ab)2 fragment, a bivalent fragment comprising two Fab fragments linked by a disulfide bridge at the hinge region; a Fd fragment consisting of the VH and CH1 domains; a Fv fragment consisting of the VL and VH domains of a single arm of an antibody; and a domain antibody (dAb) fragment (Ward et al., Nature 341:544-546, 1989), which consists of a VH domain or a VL domain. VH and VL domains can be engineered and linked together via a synthetic linker to form various types of single chain antibody designs where the VH / VL domains pair intramolecularly, or intermolecularly in those cases when the VH and VL domains are expressed by separate single chain antibody constructs, to form a monovalent antigen binding site, such as single chain Fv (scFv) or diabody, described for example in Int'l Pat. Pub. Nos. WO1998 / 44001, WO1988 / 01649, WO1994 / 13804, and WO1992 / 01047. These antibody fragments are obtained using techniques well known to those of skill in the art, and the fragments are screened for utility in the same manner as are full length antibodies.

[0044] The phrase “immunospecifically binds” refers to the ability of the disclosed antibody molecules to preferentially bind to its target (IL-5 in the case of anti-IL-5 antibody molecules) without preferentially binding other molecules in a sample containing a mixed population of molecules. Antibody molecules that immunospecifically bind IL-5 are substantially free of other antibodies having different antigenic specificities (e.g., an anti-IL-5 antibody is substantially free of antibodies that specifically bind antigens other than IL-5). Antibody molecules that immunospecifically bind IL-5, however, can have cross-reactivity to other antigens, such as orthologs of human IL-5, including Macacafascicularis (cynomolgus monkey) IL-5. The antibody molecules disclosed herein are able to immunospecifically bind both naturally-produced human IL-5 and to IL-5 which is recombinantly produced in mammalian or prokaryotic cells.

[0045] An antibody variable region consists of four “framework” regions interrupted by three “antigen binding sites.” The antigen binding sites are defined using various terms: (i) Complementarity Determining Regions (CDRs), three in the VH (HCDR1, HCDR2, HCDR3), and three in the VL (LCDRI, LCDR2, LCDR3) are based on sequence variability (Wu and Kabat J Exp Med 132:211-50, 1970; Kabat et al. Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md., 1991); and (ii) “Hypervariable regions” (“HVR” or “HV”), three in the VH (H1, H2, H3) and three in the VL (L1, L2, L3) refer to the regions of the antibody variable domains which are hypervariable in structure as defined by Chothia and Lesk (Chothia and Lesk Mol Biol 196:901-17, 1987). The AbM definition of CDRs is also widely used; it is a compromise between Kabat and Chothia numbering schemes and is so-called because it was used by Oxford Molecular's AbM antibody modelling software (Rees, A. R., Searle, S. M. J., Henry, A. H. and Pedersen, J. T. (1996) In Sternberg M. J. E. (ed.), Protein Structure Prediction. Oxford University Press, Oxford, 141-172). Other terms include “IMGT-CDRs” (Lefranc et al., Dev Comparat Immunol 27:55-77, 2003) and “Specificity Determining Residue Usage” (SDRU) (Almagro Mol Recognit 17:132-43, 2004). The International ImMunoGeneTics (IMGT) database (http: / / www_imgt_org) provides a standardized numbering and definition of antigen-binding sites. The correspondence between CDRs, HVs and IMGT delineations is described in Lefranc et al., Dev Comparat Immunol 27:55-77, 2003.

[0046] “Framework” or “framework sequences” are the remaining sequences of a variable region other than those defined to be antigen binding sites. Because the antigen binding sites can be defined by various terms as described above, the exact amino acid sequence of a framework depends on how the antigen-binding site was defined. “Human antibody,”“fully human antibody,” and like terms refers to an antibody having heavy and light chain variable regions in which both the framework and the antigen binding sites are derived from sequences of human origin. If the antibody contains a constant region, the constant region also is derived from sequences of human origin. A human antibody comprises heavy and / or light chain variable regions that are “derived from” sequences of human origin if the variable regions of the antibody are obtained from a system that uses human germline immunoglobulin or rearranged immunoglobulin genes. Such systems include human immunoglobulin gene libraries displayed on phage, and transgenic non-human animals such as mice carrying human immunoglobulin loci as described herein. “Human antibody” may contain amino acid differences when compared to the human germline or rearranged immunoglobulin sequences due to, for example, naturally occurring somatic mutations or intentional introduction of substitutions in the variable domain (framework and antigen binding sites), or constant domain. Typically, a “human antibody” is at least about 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical in amino acid sequence to an amino acid sequence encoded by a human germline or rearranged immunoglobulin gene. In some cases, a “human antibody” may contain consensus framework sequences derived from human framework sequence analyses, for example as described in Knappik et al., J Mol Biol 296:57-86, 2000, or synthetic HCDR3 incorporated into human immunoglobulin gene libraries displayed on phage, as described in, for example, Shi et al., J Mol Biol 397:385-96, 2010 and Int'l Pat. Pub. No. WO2009 / 085462. Antibodies in which antigen binding sites are derived from a non-human species are not included in the definition of “human antibody.”

[0047] Human antibodies, while derived from human immunoglobulin sequences, may be generated using systems such as phage display incorporating synthetic CDRs and / or synthetic frameworks, or can be subjected to in vitro mutagenesis to improve antibody properties in the variable regions or the constant regions or both, resulting in antibodies that do not naturally exist within the human antibody germline repertoire in vivo.

[0048] “Recombinant antibody” includes all antibodies that are prepared, expressed, created, or isolated by recombinant means, such as: antibodies isolated from an animal (e.g., a mouse) that is transgenic or transchromosomal for human immunoglobulin genes or a hybridoma prepared therefrom (described further below); antibodies isolated from a host cell transformed to express the antibody; antibodies isolated from a recombinant, combinatorial antibody library; and antibodies prepared, expressed, created, or isolated by any other means that involve splicing of human immunoglobulin gene sequences to other DNA sequences, or antibodies that are generated in vitro using Fab arm exchange.

[0049] “Monoclonal antibody” refers to a population of antibody molecules of a single molecular composition. A monoclonal antibody composition displays a single binding specificity and affinity for a particular epitope, or in a case of a bispecific monoclonal antibody, a dual binding specificity to two distinct epitopes. Monoclonal antibody therefore refers to an antibody population with single amino acid composition in each heavy and each light chain, except for possible well known alterations such as removal of C-terminal lysine from the antibody heavy chain. Monoclonal antibodies may have heterogeneous glycosylation within the antibody population. Monoclonal antibody may be monospecific or multispecific, or monovalent, bivalent or multivalent. A bispecific antibody is included in the term monoclonal antibody.

[0050] “Epitope” refers to a portion of an antigen to which an antibody specifically binds. Epitopes usually consist of chemically active (such as polar, non-polar, or hydrophobic) surface groupings of moieties such as amino acids or polysaccharide side chains and can have specific three-dimensional structural characteristics, as well as specific charge characteristics. An epitope can be composed of contiguous and / or discontiguous amino acids that form a conformational spatial unit. For a discontiguous epitope, amino acids from differing portions of the linear sequence of the antigen come in close proximity in 3-dimensional space through the folding of the protein molecule.

[0051] “Variant” refers to a polypeptide or a polynucleotide that differs from a reference polypeptide or a reference polynucleotide by one or more modifications for example, substitutions, insertions, or deletions. The term “mutation” as used herein is intended to mean one or more intentional substitutions which are made to a polypeptide or polynucleotide.

[0052] As used herein “90% identical to” encompasses at least 90% identical, 91% identical, 92% identical, 93% identical, 94% identical, 95% identical, 96% identical, 97% identical, 98% identical, 99% identical, or 100% identical to the reference item (e.g., a biological sequence). The current specification uses the term “% identical” to describe a number of sequences. As would be understood, the term “% identical” means that in a comparison of two sequences over the specified region the two sequences have the specified number of identical residues in the same position. The level of identity may be determined using CLUSTAL W with default parameters.

[0053] “Treat,”“treatment,” and like terms refer to both therapeutic treatment and prophylactic or preventative measures, and includes reducing the severity and / or frequency of symptoms, eliminating symptoms and / or the underlying cause of the symptoms, reducing the frequency or likelihood of symptoms and / or their underlying cause, improving or remediating damage caused, directly or indirectly, by the eosinophilic asthma, hypereosinophilic syndrome, nasal polyposis with eosinophilic involvement, eosinophilic granulomatosis with polyangiitis, atopic dermatitis or eosinophilic esophagitis. Treatment also includes prolonging survival as compared to the expected survival of a subject not receiving treatment. Subjects to be treated include those that have the condition or disorder as well as those prone to have the condition or disorder or those in which the condition or disorder is to be prevented.

[0054] As used herein, “administering to the subject” and similar terms indicate a procedure by which the disclosed antibody molecules or compositions comprising the same are injected into a patient such that target cells, tissues, or segments of the body of the subject are contacted with the disclosed antibody molecules.

[0055] The phrase “therapeutically effective amount” refers to an amount of the antibody molecules, as described herein, effective to achieve a particular biological or therapeutic result such as, but not limited to, biological or therapeutic results disclosed, described, or exemplified herein. The therapeutically effective amount may vary according to factors such as the disease state, age, sex, and weight of the individual, and the ability of the composition to cause a desired response in a subject. Exemplary indicators of a therapeutically effect amount include, for example, improved well-being of the patient, reduction of a disease symptom, arrested or slowed progression of disease symptoms, and / or absence of disease symptoms.

[0056] The following abbreviations are used herein: Alternaria alternata (Alternaria), Ascaris suum (A. suum); complementarity-determining region (CDR); heavy chain (HC); light chain (LC); heavy chain variable region (VH); light chain variable region (VL); surface plasmon resonance (SPR).Antibody Molecules

[0057] Disclosed herein are human antibody molecules that immunospecifically bind to human IL-5. The human antibody molecules can immunospecifically bind to human IL-5 with an equilibrium affinity constant (KD) of at least about 40 pM as determined by surface plasmon resonance (SPR). As used herein, “of at least about 40 pM” means that the disclosed antibodies immunospecifically bind human IL-5 with a KD of less than or equal to about 40 pM. For example, the disclosed antibodies can immunospecifically bind human IL-5 with a KD of about 40 pM, about 30 pM, about 20 pM, about 10 pM, or less than about 10 pM.

[0058] The disclosed human antibody molecules can comprise a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO: 6, a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO: 8, a light chain consensus CDR1 comprising the amino acid sequence of SEQ ID NO: 1, a light chain consensus CDR2 comprising the amino acid sequence of SEQ ID NO: 2, and a light chain consensus CDR3 comprising the amino acid sequence of SEQ ID NO: 3.

[0059] The light chain consensus CDR1 comprises the amino acid sequence of GX1X2X3X4X5X6KX7X8Y (SEQ ID NO: 1), wherein:

[0060] X1 is G or K;

[0061] X2 is N or D;

[0062] X3 is N or H;

[0063] X4 is I or A;

[0064] X5 is G or D;

[0065] X6 is S or K;

[0066] X7 is N or H; and

[0067] X8 is V or A.

[0068] In some embodiments, the light chain CDR1 amino acid sequence can comprise GGNNIGSKNVY (SEQ ID NO: 5). In some embodiments, the light chain CDR1 amino acid sequence can comprise GKNNIGSKNVY (SEQ ID NO: 21). In some embodiments, the light chain CDR1 amino acid sequence can comprise GGDNIGSKNVY (SEQ ID NO: 24). In some embodiments, the light chain CDR1 amino acid sequence can comprise GGNHIGSKNVY (SEQ ID NO: 27). In some embodiments, the light chain CDR1 amino acid sequence can comprise GGNNAGSKNVY (SEQ ID NO: 30). In some embodiments, the light chain CDR1 amino acid sequence can comprise GGNNIDSKNVY (SEQ ID NO: 66). In some embodiments, the light chain CDR1 amino acid sequence can comprise GGNNIGKKNVY (SEQ ID NO: 33). In some embodiments, the light chain CDR1 amino acid sequence can comprise GGNNIGSKHVY (SEQ ID NO: 36). In some embodiments, the light chain CDR1 amino acid sequence can comprise GGNNIGSKNAY (SEQ ID NO: 39).

[0069] The light chain CDR2 amino acid sequence comprises DDX8X9RPS (SEQ ID NO: 2), wherein:

[0070] X8 is S or L; and

[0071] X9 is D or S.

[0072] In some embodiments, the light chain CDR2 amino acid sequence can comprise DDSDRPS (SEQ ID NO: 7). In some embodiments, the light chain CDR2 amino acid sequence can comprise DDLDRPS (SEQ ID NO: 42). In some embodiments, the light chain CDR2 amino acid sequence can comprise DDSSRPS (SEQ ID NO: 45).

[0073] The light chain CDR3 amino acid sequence comprises QVWX10SSSDX11VX12 (SEQ ID NO: 3), wherein:

[0074] X10 is D or L;

[0075] X11 is H, S, Y, or D; and

[0076] X12 is V, A, or W.

[0077] In some embodiments, the light chain CDR3 amino acid sequence can comprise QVWDSSSDHVV (SEQ ID NO: 15). In some embodiments, the light chain CDR3 amino acid sequence can comprise QVWLSSSDHVV (SEQ ID NO: 48). In some embodiments, the light chain CDR3 amino acid sequence can comprise QVWDSSSDSVV (SEQ ID NO: 51). In some embodiments, the light chain CDR3 amino acid sequence can comprise QVWDSSSDYVV (SEQ ID NO: 54). In some embodiments, the light chain CDR3 amino acid sequence can comprise QVWDSSSDDVV (SEQ ID NO: 57). In some embodiments, the light chain CDR3 amino acid sequence can comprise QVWDSSSDHVA (SEQ ID NO: 60). In some embodiments, the light chain CDR3 amino acid sequence can comprise QVWDSSSDHVW (SEQ ID NO: 63).

[0078] The disclosed human antibody molecules can comprise a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO: 6, a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO: 8, a light chain CDR1 comprising the amino acid sequence of SEQ ID NOs: 5, 21, 24, 27, 30, 33, 36, 39, or 66, a light chain CDR2 comprising the amino acid sequence of SEQ ID NOs: 7, 42, or 45, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NOs: 15, 48, 51, 54, 57, 60, or 63. Exemplary antibody molecules comprise a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO: 6, a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO: 8, and

[0079] a. a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 5, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 15;

[0080] b. a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 21, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 15;

[0081] c. a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 24, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 15;

[0082] d. a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 27, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 15;

[0083] e. a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 30, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 15;

[0084] f. a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 33, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 15;

[0085] g. a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 36, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 15;

[0086] h. a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 39, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 15;

[0087] i. a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 66, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 15;

[0088] j. a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 5, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 42, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 15;

[0089] k. a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 5, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 45, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 15;

[0090] l. a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 5, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 48;

[0091] m. a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 5, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 51;

[0092] n. a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 5, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 54;

[0093] o. a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 5, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 57;

[0094] p. a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 5, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 60; or

[0095] q. a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 5, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 63;

[0096] wherein the position of the amino acid residues of the CDR is determined according to AbM.

[0097] The disclosed antibody molecules can comprise a heavy chain variable region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 16 and a light chain variable region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 17, 22, 25, 28, 31, 34, 37, 40, 43, 46, 49, 52, 55, 58, 61, 64, or 67, wherein the variability (i.e. the at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identity) occurs outside of the CDR sequence. Exemplary antibody molecules are provided in Table 1 and Table 15.

[0098] TABLE 1Antibody chain / domain composition summaryAntibodyVH protein / VL protein / HC protein / LC protein / ID(SEQ ID)(SEQ ID)(SEQ ID)(SEQ ID)3A53A5 VH3A5 VL3A5 HC3A5 LC(SEQ ID NO: 10)(SEQ ID NO: 11)(SEQ ID NO: 12)(SEQ ID NO: 13)3A5.0013A5 VH3A5 VL3A5.001 HC3A5 LC(SEQ ID NO: 10)(SEQ ID NO: 11)(SEQ ID NO: 14)(SEQ ID NO: 13)3A5.0403A5.040 VH3A5.040 VL3A5.040 HC3A5.040 LC(SEQ ID NO: 16)(SEQ ID NO: 17)(SEQ ID NO: 18)(SEQ ID NO: 19)3A5.0463A5.040 VH3A5.040 VL3A5.046 HC3A5.040 LC(SEQ ID NO: 16)(SEQ ID NO: 17)(SEQ ID NO: 20)(SEQ ID NO: 19)3A5.0633A5.040 VH3A5.040 VL +3A5.040 HC3A5.040 LC +(SEQ ID NO: 16)G25K(SEQ ID NO: 18)G25K(SEQ ID NO: 22)(SEQ ID NO: 23)3A5.0703A5.040 VH3A5.040 VL +3A5.040 HC3A5.040 LC +(SEQ ID NO: 16)N26D(SEQ ID NO: 18)N26D(SEQ ID NO: 25)(SEQ ID NO: 26)3A5.0823A5.040 VH3A5.040 VL +3A5.040 HC3A5.040 LC +(SEQ ID NO: 16)N27H(SEQ ID NO: 18)N27H(SEQ ID NO: 28)(SEQ ID NO: 29)3A5.0843A5.040 VH3A5.040 VL +3A5.040 HC3A5.040 LC +(SEQ ID NO: 16)I28A(SEQ ID NO: 18)I28A(SEQ ID NO: 31)(SEQ ID NO: 32)3A5.0973A5.040 VH3A5.040 VL +3A5.040 HC3A5.040 LC +(SEQ ID NO: 16)G29D(SEQ ID NO: 18)G29D(SEQ ID NO: 67)(SEQ ID NO: 68)3A5.1073A5.040 VH3A5.040 VL +3A5.040 HC3A5.040 LC +(SEQ ID NO: 16)S30K(SEQ ID NO: 18)S30K(SEQ ID NO: 34)(SEQ ID NO: 35)3A5.1253A5.040 VH3A5.040 VL +3A5.040 HC3A5.040 LC +(SEQ ID NO: 16)N32H(SEQ ID NO: 18)N32H(SEQ ID NO: 37)(SEQ ID NO: 38)3A5.1273A5.040 VH3A5.040 VL +3A5.040 HC3A5.040 LC +(SEQ ID NO: 16)V33A(SEQ ID NO: 18)V33A(SEQ ID NO: 40)(SEQ ID NO: 41)3A5.1613A5.040 VH3A5.040 VL +3A5.040 HC3A5.040 LC +(SEQ ID NO: 16)S52L(SEQ ID NO: 18)S52L(SEQ ID NO: 43)(SEQ ID NO: 44)3A5.1693A5.040 VH3A5.040 VL +3A5.040 HC3A5.040 LC +(SEQ ID NO: 16)D53S(SEQ ID NO: 18)D53S(SEQ ID NO: 46)(SEQ ID NO: 47)3A5.2323A5.040 VH3A5.040 VL +3A5.040 HC3A5.040 LC +(SEQ ID NO: 16)D92L(SEQ ID NO: 18)D92L(SEQ ID NO: 49)(SEQ ID NO: 50)3A5.276*3A5.040 VH3A5.040 VL +3A5.040 HC3A5.040 LC +(SEQ ID NO: 16)H95bS(SEQ ID NO: 18)H95bS(SEQ ID NO: 52)(SEQ ID NO: 53)3A5.278*3A5.040 VH3A5.040 VL +3A5.040 HC3A5.040 LC +(SEQ ID NO: 16)H95bY(SEQ ID NO: 18)H95bY(SEQ ID NO: 55)(SEQ ID NO: 56)3A5.279*3A5.040 VH3A5.040 VL +3A5.040 HC3A5.040 LC +(SEQ ID NO: 16)H95bD(SEQ ID NO: 18)H95bD(SEQ ID NO: 58)(SEQ ID NO: 59)3A5.2943A5.040 VH3A5.040 VL +3A5.040 HC3A5.040 LC +(SEQ ID NO: 16)V97A(SEQ ID NO: 18)V97A(SEQ ID NO: 61)(SEQ ID NO: 62)3A5.3023A5.040 VH3A5.040 VL +3A5.040 HC3A5.040 LC +(SEQ ID NO: 16)V97W(SEQ ID NO: 18)V97W(SEQ ID NO: 64)(SEQ ID NO: 65)*The lower case “b” in each of these sequences refers to the Kabat numbering for the CDR position. Kabat numbering allows for CDRs of variable sizes, by using alphanumeric numbering to denote amino acid insertions at certain positions. In these CDR sequences, additional amino acids were present, which were numbered as positions 95a and 95b (corresponding to Kabat positions 95A and 95B respectively). Thus, for antibody 3A5.276, for example, H95bS indicates a histidine (“H”) to serine (“S”) mutation at position 95B according to Kabat number relative to the 3A5.040 VL chain. The lower case notation is therefore used here to distinguish the Kabat number, separate from the mutation at the indicated Kabat position.

[0099] In some embodiments, the disclosed antibody molecules can comprise a heavy chain variable region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 16 and

[0100] a. a light chain variable region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 17;

[0101] b. a light chain variable region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 22;

[0102] c. a light chain variable region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 25;

[0103] d. a light chain variable region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 28;

[0104] e. a light chain variable region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 31;

[0105] f. a light chain variable region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 34;

[0106] g. a light chain variable region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 37;

[0107] h. a light chain variable region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 40;

[0108] i. a light chain variable region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 43;

[0109] j. a light chain variable region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 46;

[0110] k. a light chain variable region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 49;

[0111] l. a light chain variable region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 52;

[0112] m. a light chain variable region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 55;

[0113] n. a light chain variable region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 58;

[0114] o. a light chain variable region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 61;

[0115] p. a light chain variable region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 64; or

[0116] q. a light chain variable region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 67,

[0117] wherein the variability (i.e. the at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identity) occurs outside of the CDR sequence.

[0118] The disclosed antibody molecules can comprise one or more mutations, deletions, or insertions, in the framework and / or constant regions. In some embodiments, an IgG4 antibody molecule can comprise a S228P mutation. S228 is located in the hinge region of the IgG4 antibody molecule. Mutation of the serine (“S”) to a proline (“P”) serves to stabilize the hinge of the IgG4 and prevent Fab arm exchange in vitro and in vivo. In some embodiments, the antibody molecules can comprise one or more modifications which increase the in vivo half life of the antibody molecules. For instance in certain embodiments the antibody can comprise a M252Y mutation, a S254T mutation, and a T256E mutation (collectively referred to as the “YTE” mutation). M252, S254, and T256 are located in in the CH2 domain of the heavy chain. Mutation of these residues to tyrosine (“Y”), threonine (“T”), and glutamate (“E”), respectively, protects the antibody molecules from lysosomal degradation, thereby enhancing the serum half-life of the antibody molecules. Based on the example of other antibodies, it is contemplated that the introduction of the YTE mutation in an anti-IL-5 antibody may provide sufficient extension of serum half life to allow for administration regimes with 3 months or longer inter-dosing intervals. In some embodiments, the antibody molecules can comprise a deletion of the heavy chain C-terminal lysine residue. Deletion of the heavy chain C-terminal lysine residue reduces heterogeneity of the antibody molecules when produced by mammalian cells. In some embodiments, the antibody molecules can comprise a combination of mutations, deletions, or insertions. For example, in some aspects, the disclosed antibody molecules can comprise a S228P mutation and a deletion of a heavy chain C-terminal lysine residue. The disclosed antibodies comprising a heavy chain sequence of SEQ ID NO: 18, for example, comprise a S228P mutation and a deletion of a heavy chain C-terminal lysine residue. In some aspects, the disclosed antibody molecule can comprise a S228P mutation, a M252Y mutation, a S254T mutation, a T256E mutation, and a deletion of a heavy chain C-terminal lysine residue. The 3A5.046 antibody, for example, which comprises a heavy chain of SEQ ID NO: 20, comprises a S228P mutation, a M252Y mutation, a S254T mutation, a T256E mutation, and a deletion of a heavy chain C-terminal lysine residue.

[0119] The antibody molecule can comprise an IgG1 or IgG4 heavy chain constant region and a lambda light chain constant region. In some embodiments, the antibody molecule comprises an IgG1 heavy chain constant region and a lambda light chain constant region (antibody 3A5, for example). In some embodiments, the antibody molecule comprises an IgG4 heavy chain constant region and a lambda light chain constant region.

[0120] The disclosed antibody molecules can comprise a heavy chain comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NOs: 18 or 20 and a light chain comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 19, 23, 26, 29, 32, 35, 38, 41, 44, 47, 50, 53, 56, 59, 62, 65, or 68, wherein the variability (i.e. the at least 90% identity) occurs outside of the CDR sequence. Exemplary antibody molecules are provided in Table 1 and Table 15. In some embodiments, the antibody molecules can comprise a heavy chain comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 18 and

[0121] a. a light chain comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 19;

[0122] b. a light chain comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 23;

[0123] c. a light chain comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 26;

[0124] d. a light chain comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 29;

[0125] e. a light chain comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 32;

[0126] f. a light chain comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 35;

[0127] g. a light chain comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 38;

[0128] h. a light chain comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 41;

[0129] i. a light chain comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 44;

[0130] j. a light chain comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 47;

[0131] k. a light chain comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 50;

[0132] l. a light chain comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 53;

[0133] m. a light chain comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 56;

[0134] n. a light chain comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 59;

[0135] o. a light chain comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 62;

[0136] p. a light chain comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 65; or

[0137] q. a light chain comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 68,

[0138] wherein the variability (i.e. the at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identity) occurs outside of the CDR sequence.

[0139] The disclosed antibody molecules can comprise a heavy chain comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 20 and a light chain comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 19, wherein the variability (i.e. the at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identity) occurs outside of the CDR sequence.

[0140] In some embodiments, the antibody molecules are full length antibody molecules (with or without a deletion of the heavy chain C-terminal lysine residue). In other embodiments, the antibody molecules are antigen binding fragments. Suitable antibody binding fragments include, but are not limited to, a Fab fragment, a Fab2 fragment, or a single chain antibody.

[0141] The antibody molecules can have one or more of the following properties:

[0142] a. binds to human IL-5 with an equilibrium affinity constant (KD) of at least about 40 pM as determined by surface plasmon resonance;

[0143] b. reduces binding of IL-5 to the IL-5 receptor;

[0144] c. has a serum half-life of at least about 20 days; or

[0145] d. binds human and cynomolgus monkey IL-5 but not mouse, rat, or guinea pig IL-5.

[0146] Pharmaceutical compositions comprising any of the disclosed antibody molecules are also provided.

[0147] Also provided are nucleic acid molecules encoding any of the disclosed antibody molecules and vectors comprising the disclosed nucleic acid molecules.

[0148] Cells transformed to express any of the disclosed antibody molecules are further provided.Methods and Uses

[0149] The disclosed antibody molecules, or pharmaceutical compositions comprising the same, can be used to treat eosinophilic asthma, hypereosinophilic syndrome, nasal polyposis with eosinophilic involvement, eosinophilic granulomatosis with polyangiitis, atopic dermatitis and eosinophilic esophagitis. Any of the antibody molecule characteristics disclosed herein apply equally to the antibodies used in the disclosed methods and uses.

[0150] Disclosed herein are methods of treating a subject having eosinophilic asthma, hypereosinophilic syndrome, nasal polyposis with eosinophilic involvement, eosinophilic granulomatosis with polyangiitis, atopic dermatitis or eosinophilic esophagitis comprising administering to the subject a therapeutically effective amount of any of the antibody molecules disclosed herein, or pharmaceutical compositions comprising the same, to treat the eosinophilic asthma, hypereosinophilic syndrome, nasal polyposis with eosinophilic involvement, eosinophilic granulomatosis with polyangiitis, atopic dermatitis or eosinophilic esophagitis.

[0151] The use of an effective amount of any of the disclosed antibody molecules, or pharmaceutical compositions comprising the same, in the treatment of eosinophilic asthma, hypereosinophilic syndrome, nasal polyposis with eosinophilic involvement, eosinophilic granulomatosis with polyangiitis, atopic dermatitis or eosinophilic esophagitis is also provided.

[0152] Also provided is the use of any of the disclosed antibody molecules, or pharmaceutical compositions comprising the same, in the manufacture of a medicament for the treatment of eosinophilic asthma, hypereosinophilic syndrome, nasal polyposis with eosinophilic involvement, eosinophilic granulomatosis with polyangiitis, atopic dermatitis or eosinophilic esophagitis.EXAMPLES

[0153] The following examples are provided to further describe some of the embodiments disclosed herein. The examples are intended to illustrate, not to limit, the disclosed embodiments.Generation of Anti-Human-IL-5 Antibodies

[0154] Anti-human-IL-5 antibodies were obtained from transgenic rats (OMT) with human V-genes cloned into their genomes and which produce antibodies with human V-domains and rat Fc domains. Briefly, the transgenic rats were genetically immunized with DNA encoding IL-5 four times over 21 days (on days 0, 7, 14, 21) and boosted with recombinant human IL-5 at day 28 of the immunization protocol. Serum antibody titres were determined at days 0 and 38 of the immunization protocol by an ELISA assay using recombinant human IL-5. Briefly, sera from each animal were diluted in PBS 1% BSA and were tested using ELISA plates coated with 1 μg / ml human IL-5, or BSA as a control. A goat anti-rat IgG R-phycoerythrin conjugate (SOUTHERN BIOTECH®, #3030-09) was used at 10 μg / ml as a secondary antibody. Specific animals were chosen for hybridoma fusion based on these serum titres.

[0155] To generate hybridomas which produce monoclonal antibodies to human IL-5, splenocytes and / or lymph node cells from immunized animals were isolated and fused to P3X63Ag8.653 non-secreting mouse myeloma cells (ATCC®, CRL-1580). Cells were plated at approximately 1×105 cells / mL in flat bottom microtiter plates, followed by a two week incubation in selective medium which included 10% fetal clone serum and 1×HAT (SIGMA). Hybridomas were expanded by serial passage through four media changes in 96-well plates (96-well stages 1 to 4), then into T25 and finally T75 flasks.

[0156] The supernatants from hybridoma clones were assayed during the hybridoma expansion process, initially in a whole-cell ELISA using cells transfected with GPI-anchored human IL-5 and then ELISA using a recombinant human IL-5 ELISA assay (the latter as described above). Hybridomas which survived the scale-up process to T75 stage and which gave signals above a given threshold for binding in both of these assays were frozen as cell pellets, for cloning and sequencing of Ig v-domains. Approximately 20 hybridomas producing chimeric IgGs were selected for cloning following these screening steps.

[0157] The human v-domains from the candidate chimeric IgGs were isolated by generation of cDNA from hybridoma cell pellets, PCR amplification of v-domains, subcloning, and DNA sequencing. A total of approximately 35 heavy and light chain combinations were obtained from the sequencing of these hybridomas. All antibodies were cloned into a mammalian expression vector and transiently transfected into HEK-293 cells. Antibodies were purified using standard Protein A purification protocols.Functional Testing and Characterization of Antibodies

[0158] Selected antibodies in human IgG1 format were first assayed for specificity in a human IL-5 ELISA. Briefly, antibodies were diluted in PBS 0.1% BSA and were tested using ELISA plates coated with 1 μg / ml human IL-5, or an irrelevant control protein. A horseradish peroxidase-conjugated Anti-IgG antibody was used as a secondary antibody.

[0159] Antibodies that showed immunospecific binding to human IL-5 were ranked for potency in a human IL-5-dependent cell proliferation assay using human TF-1.6G4 (a derivative of the human erythroleukemic cell line TF-1) cells. The TF-1.6G4 cell line was subcloned and selected for enhanced surface expression of IL-5Rα and consistent proliferative response to human IL-5. The TF-1.6G4 cell line was maintained in culture following standard conditions used for the TF-1 human erythroleukemic cell line (ATCC®: CRL-2003). Briefly, dilutions of each antibody were incubated in the presence of 45 pM human IL-5 and 5×104 TF-1.6G4 cells per well, incubated for 48 h, and cell proliferation was determined using the CELLTITER-GLO® Luminescent Cell Viability Assay (PROMEGA®, WI). All proliferation and inhibition curves were fitted using a three- or four-parameter dose-response model in GraphPad Prism®6 (Version 6.04) software. Table 2 summarizes these results.

[0160] TABLE 2Summary of screening and characterizationof an initial test panel in IgG1 format.EquilibriumInhibitsSpecificityTestIC50 (pM)AffinityIL-5RαconfirmedAntibodyin TF1.6G4(KD)bindingby ELISA3A5 35.0736 pMYesYes1A3 55.518 pMYesYes2B4 56.7542 pMYesYes6G10 72.3461 pMYesYes1E8 72.4528 pMYesYes3G4 80.9241 pMYesYes6C9 81.8941 pMYesYes5H1 86.4236 pMYesYes1H10 93.2618 pMYesYes5H11109.336 pMNoYes5G9184.452 pMYesYes1B2214.445 pMYesYes

[0161] In Table 2, the results of tests were ranked in order of TF1.6G4 assay potency. The initial test panel was selected based upon potency, affinity, IL-5Rα inhibition and sequence liabilities (immunogenicity and developability).

[0162] BIACORE™ assays were used to determine the affinity of test antibodies to recombinant human IL-5 and their potency in inhibiting IL-5Rα binding to human IL-5. Table 2 summarizes these results. Binding affinity of test antibodies to human IL-5 (KD; FIG. 6) was determined on a BIACORE™ T200 system (GE HEALTHCARE®) by coating a BIACORE™ Series S Sensor Chip Protein A (GE HEALTHCARE®) with selected purified IgG antibody to a capture level of 75 RU, then recombinant human IL-5 was injected at 60 μL / min across a 7-step two-fold serial dilution range, starting at 1 μg / mL. All experiments were run using HBS-EP+ buffer (GE HEALTHCARER). The resulting sensorgrams were double-referenced (test flowcell values subtracted from control (Protein A surface with no coated antibody) flowcell values and also a buffer blank). Binding constants were determined by fitting a 1:1 Langmuir binding model to double-referenced sensorgrams.

[0163] To determine whether each test antibody inhibited the binding of IL-5 to IL-5Rα, either a BIACORE™ T200 or a BIACORE™ 3000 system (GE HEALTHCARE®) was used. A BIACORE™ CM5 Sensor Chip was first derivatized with a Fab capture kit (GE HEALTHCARER) in accordance with the manufacturer's instructions on two adjacent (test and control) flowcells. This surface was used to capture each purified IgG test antibody on a single test flowcell. Recombinant human IL-5 at 5 μg / mL or a buffer blank was then injected across both test and control flowcells, to saturate the test flowcell surface and control for non-specific association of IL-5, respectively. A second injection of purified IgG test antibody at 10 μg / mL or a buffer blank was performed on the test flowcell to block free IL-5 binding sites and control for dissociation of the IgG test antibody from the Fab capture antibody, respectively.

[0164] Subsequent injection of IL-5Rα-Fc (R&D SYSTEMS®) at either 5 or 20 μg / mL or a buffer blank across both flowcells was used to determine whether the IgG test antibody blocked the interaction between IL-5 and IL-5Rα or to control for dissociation of IgG antibody from the Fab capture antibody during this step. Antibodies which inhibited the binding of IL-5 to IL-5Rα showed a markedly reduced signal upon injection of IL-5Rα (Table 2). A triple reference subtraction method was used to analyze data in the manner detailed above. All data was exported from BIACORE™ evaluation software and subtracted in Excel (MICROSOFT®) software. All experiments were run using HBS-EP+ buffer (GE HEALTHCARE®).

[0165] A smaller panel of test antibodies (3A5, 5H11 and 2B4 in Table 2) was chosen on the basis of these results. The antibodies were reformatted as human IgG4 and tested in the same assays. The IgG4 version of antibody 3A5 (originally an IgG1) was designated as 3A5.001. This fully-human antibody was demonstrated to have equivalent potency to the original test 3A5 in IgG1 format (FIG. 1).

[0166] A variant of antibody 3A5.001 (designated 3A5.040) was prepared with specific amino acid substitutions (VH: S

[68] T, N[82A]S; VL: S[2]Y, I[3]V, Y

[92] D, wherein the residues in square brackets represent the Kabat positions) introduced in the V-domain regions to remove predicted T-cell epitopes. Antibody 3A5.040 was demonstrated to have equivalent potency to its parental antibody, 3A5.001 (FIG. 2) in the TF-1.6G4 assay.CDR Scanning of Antibody 3A5.040

[0167] Generation of antibody 3A5.040 variants—Single-mutant variants of antibody 3A5.040 were made by substituting one of a group of nine representative amino acids—A, S, L, Y, D, Q, K, H, W—at each amino acid position in the light chain CDR1 (CDR-L1), the light chain CDR2 (CDR-L2), the heavy chain CDR1 (CDR-H1) and the heavy chain CDR2 (CDR-H2) (as defined by AbM nomenclature). Antibody variants were also made by substituting one of a group of ten representative amino acids—A, S, L, Y, D, Q, K, H, W, P—at each CDR amino acid position in the light chain CDR3 (CDR-L3), heavy chain CDR3 (CDR-H3), and at Kabat positions 93 and 94 in the variable heavy chain. A complete list of all single-mutant antibody variants generated is shown in FIG. 3 (variable heavy chain) and FIG. 4 (variable light chain), respectively.

[0168] Construction of Vectors Expressing Antibodies—Variable region variants were generated by back-translation of amino acid sequences into DNA sequences which were subsequently synthesized de novo by assembly of synthetic oligonucleotides. Variable heavy (VH) variants were subcloned into a mammalian expression vector containing a human constant region to produce full-length antibody heavy chains (human IgG4 heavy chain CHI, hinge, CH2, and CH3 domains). Similarly, variable light (VL) variants were subcloned into a mammalian expression vector containing a human lambda light chain constant region to produce full-length antibody lambda chains.

[0169] Expression of antibody variants—Antibodies were produced by co-transfecting separate expression vectors encoding antibody heavy chains and light chains into EXPI293® cells (LIFE TECHNOLOGIES®, Carlsbad, CA). Each single-mutant chain was paired with a parental chain for protein expression in the EXP1293® system. For each 20 mL transfection, 3.6×107 cells were required in 20 mL of EXP1293® Expression Medium. On the day prior to transfection, cells were seeded at a density of 0.9×106 viable cells / mL and incubated overnight at 37° C. in a humidified atmosphere of 8% CO2 in air on an orbital shaker rotating at 200 rpm. On the day of transfection, the cell number and viability were determined using an automated cell counter. Only cultures with >98% viable cells were used. For each 20 mL transfection, lipid-DNA complexes were prepared by diluting 10 μg of heavy chain DNA and 10 μg of light chain DNA in OPTI-MEM® (LIFE TECHNOLOGIES®, Carlsbad, CA) I Reduced Serum Medium (Cat. no. 31985-062) to a total volume of 1.0 mL. 54 μL of EXPIFECTAMINE® 293 Reagent (LIFE TECHNOLOGIES®, Carlsbad, CA) was diluted in OPTI-MEM® I medium to a total volume of 1.0 mL. Both vials were mixed gently and incubated for 5 minutes at room temperature. Following incubation, the diluted DNA was mixed with the diluted EXPIFECTAMINE® 293 Reagent and the DNA-EXPIFECTAMINE® 293 Reagent mixture and incubated a further 20 minutes at room temperature to allow the formation of DNA-EXPIFECTAMINE® 293 Reagent complexes. Following incubation, 2 mL of DNA-EXPIFECTAMINE® 293 Reagent complex was added to each 50 mL bioreactor tube (TPP TECHNO PLASTIC PRODUCTS® AG). 2 mL of OPTI-MEM® (LIFE TECHNOLOGIES®, Carlsbad, CA) I medium was added to the negative control tube instead of DNA-EXPIFECTAMINE® 293 Reagent complex. The cells were incubated in a 37° C. incubator with a humidified atmosphere of 8% CO2 in air on an orbital shaker rotating at 200 rpm. Approximately 16-18 hours post-transfection, 100 μL of EXPIFECTAMINE® 293 Transfection Enhancer 1 and 1.0 mL of EXPIFECTAMINE® 293 Transfection Enhancer 2 were added to each bioreactor. Supernatants were harvested at approximately 48 hours post-transfection.

[0170] Purification of antibody variants—Each antibody variant was expressed in EXP1293® cells in either 20 or 100 mL of cell culture. Cultures were spun down in 50 mL falcon tubes at 3000×g for 20 minutes, and supernatants were filtered using a 0.22 μm filter. Supernatants were purified using a GILSON® ASPEC GX274 robot. Briefly, SPE cartridges (AGILENT®, 12131014) packed with 1.2 mL MABSELECT SURE® protein A resin (GE HEALTHCARE®) were pre-equilibrated with 3 column volumes of 1×PBS. Supernatant was run over the columns followed by a 4 mL 1×PBS wash. Each column was washed with 9 mL of 1 M citric acid, pH 2.9. Antibodies were eluted with 2 mL 0.1 M citric acid, pH 2.9. Antibodies were desalted into Sorensens PBS (5 mM KH2PO4, 3 mM Na2HPO4·2H2O, 145.4 mM NaCl (pH˜5.8)) using PD-10 columns (GE HEALTHCARE®).

[0171] Determination of 3A5.040 variant antibody titre in supernatant by BIACORE™—Supernatants containing antibody from expression in EXPI293® cells were analyzed using a Protein A series S chip on the BIACORE™ T200 to determine their titre and rank them with respect to the parental antibody 3A5.040. Each supernatant sample was diluted with running buffer (1×HBS-EP+, 350 mM NaCl) and captured by injection at 60 μL / min onto flow cell (FC) 2. The resulting capture level in response units (RUs) were measured on FC 2-1 by subtracting a report point 5 sec after cycle start from one 5 sec after injection. The surface was regenerated by injecting 50 mM NaOH for 12 sec at a flow rate of 60 μL / min onto FC 1 and 2 every 4 cycles. After each regeneration, the surface and sensorgram was stabilized for 120 sec by injection of running buffer. Multiple batches were run due to the large number of antibodies that were screened in this manner.

[0172] The capture levels (in BIACORE™ response units “RU”) obtained for each supernatant sample were adjusted by multiplying the values obtained for each 30 sec injection by the supernatant dilution factor, enabling comparison of capture levels between supernatants diluted to varying degrees across different experimental runs. The relative antibody expression level (titre) of each mutant was compared to a batch-specific 3A5.040 supernatant, (Table 3) using the following formula:(adjusted 3A5.040 reference RU for 30 sec injection / adjusted variant antibody RU for 30 sec injection)×100=proportional titre of parental antibody(% 3A5.040 titre)   Formula 1

[0173] Variant antibodies with a higher titre than parent 3A5.040 antibody had a “% 3A5.040 titre” value >100% and those with a lower titre, a value <100%. This enabled the identification of variant antibodies that might have improved expression over the parental antibody, 3A5.040. The titres of some variants were not determined by BIACORE™ analysis of supernatant samples, but were expressed and purified as above and their purified yields determined by spectrophotometric analysis (A280), then compared to parental 3A5.040 antibody (Table 5).

[0174] Determination of 3A5.040 variant antibody IL-5-binding kinetics by BIACORE™—Supernatants containing antibody from expression in EXPI293® cells or purified antibodies were analyzed using a Protein A series S chip on a BIACORE™ T200 system (GE HEALTHCARE®) to determine their binding affinity for recombinant human IL-5 and rank them with respect to the parental antibody 3A5.040.

[0175] Each antibody was diluted in running buffer (1×HBS-EP+, 350 mM NaCl) and captured at 60 μL / min to approximately 50 RU on FC 2. The surface and sensorgram was then stabilized for 120 sec by injection of running buffer. Recombinant human IL-5 at 5 μg / mL or running buffer was injected onto FC 1 and 2 at a flow rate of 60 μL / min for 70 sec. The IL-5 was allowed to dissociate in running buffer for 300 sec. The surface was regenerated by injecting 50 mM NaOH for 12 sec at a flow rate of 60 μL / min onto FC 1 and 2. Buffer was injected to further clean up drift for 60 sec at a flow rate of 60 μL / min onto FC 1 and 2. The surface was stabilized for 300 sec with running buffer over FC 1 and 2. Supernatant containing the parental 3A5.040 antibody and that had been transfected with each batch was run approximately every 25 cycles throughout each run. A purified sample of 3A5.040 and an IgG4 lambda isotype control was run with every batch as a measure of inter-assay variability. Multiple batches were run due to the large number of antibodies that were screened in this manner.

[0176] Data was double-referenced (flow cell 2 was subtracted from flow cell 1 and a buffer blank) and sensorgrams fitted to a 1:1 Langmuir binding model with BIACORE™ Evaluation software. A kd (off-rate) value was calculated for each 3A5.040 supernatant sample throughout the run and these were averaged to give a reference kd, with the exception of run 2.1, which only had one 3A5.040 supernatant sample (Table 3 and Tables 6-11). The calculated kd value for each supernatant antibody was compared against the 3A5.040 average reference kd using the following formula:(3A5.040 reference kd / variant antibody kd)×100=proportional kd of parental antibody(% 3A5.040 kd)   Formula 2

[0177] Variant antibodies with a lower kd (slower off-rate) than parent 3A5.040 antibody had a “% 3A5.040 kd” value >100% and those with a higher kd (faster off-rate) a value <100%. This enabled the identification of variant antibodies that might have improved binding kinetics for IL-5 and therefore improved function over the parental antibody, 3A5.040. For variants where only purified protein was made (Table 6), the BIACORE™ kinetic analysis was performed in triplicate as above using these purified antibodies and the average values from each triplicate analysis used to perform the proportional kd calculation, as above (Table 4 and Tables 8-11).

[0178] TABLE 3Determination of supematant 3A5.040 variant antibody titre and IL-5 binding kinetics by Biacore%%RunAntibodyReplicateAmino acidCaptureRmax3A5.0403A5.04#IDNumberSubstitutionlevelka (1 / Ms)kd (1 / s)KD (M)(RU)kd0 titre1.13A5.0401Wild type51.67.20E+051.15E−041.60E−1016.31.13A5.0402Wild type51.67.27E+058.96E−051.23E−1016.61.13A5.0403Wild type51.87.18E+059.21E−051.28E−1016.71.13A5.0404Wild type526.96E+059.95E−051.43E−1016.81.13A5.040AverageWild type51.757.15E+059.91E−051.39E−1016.61001001.1IgG4 54.4N / AN / AN / A0.7N / AN / AIsotype1.13A5.040Wild type56.47.17E+051.06E−041.48E−1017.893.0(Purified)1.13A5.304VH G26A54.67.41E+057.73E−051.04E−1017.1128491.13A5.305VH G26S53.17.36E+051.09E−041.48E−1016.791591.13A5.306VH G26L49.17.49E+058.74E−051.17E−1015.6113151.13A5.308VH G26D52.56.75E+058.57E−051.27E−1016.4116611.13A5.309VH G26Q55.17.47E+059.23E−051.24E−1017.4107861.13A5.310VH G26K51.87.36E+051.01E−041.37E−1016.4981001.13A5.311VH G26H50.97.60E+051.11E−041.46E−1016.389851.13A5.312VH G26W56.18.54E+051.21E−041.41E−1018821071.13A5.313VH G27A54.56.97E+051.17E−041.68E−1017.385991.13A5.314VH G27S50.97.16E+059.81E−051.37E−1016.2101951.13A5.315VH G27L54.37.01E+051.31E−041.87E−1017.1761181.13A5.316VH G27Y566.66E+058.56E−041.29E−0916.912801.13A5.317VH G27D57.46.56E+051.16E−041.76E−1017.885731.13A5.318VH G27Q55.27.31E+051.49E−042.04E−1017.467821.13A5.319VH G27K54.87.70E+052.04E−042.65E−1017.549951.13A5.320VH G27H54.47.12E+052.43E−043.42E−1016.941711.13A5.322VH S28A587.49E+051.05E−041.41E−101894621.13A5.323VH S28L53.87.18E+059.30E−051.30E−1017107701.13A5.324VH S28Y54.88.13E+058.60E−051.06E−1017115511.13A5.325VH S28D54.96.03E+059.16E−051.52E−1017108681.13A5.326VH S28Q56.96.78E+059.97E−051.47E−1017.799771.13A5.327VH S28K53.77.22E+051.15E−041.59E−1016.986841.13A5.328VH S28H547.82E+051.23E−041.57E−1016.981711.13A5.329VH S28W558.33E+051.36E−041.63E−101773411.13A5.330VH I29A56.36.54E+051.69E−042.58E−1016.859301.13A5.331VH I29S53.35.87E+052.04E−043.48E−1015.749281.13A5.332VH I29L55.38.26E+051.16E−041.41E−1017.585751.13A5.333VH I29Y55.45.44E+059.31E−041.71E−0915.711241.13A5.334VH I29D52.83.53E+065.37E−031.52E−0911.92211.13A5.336VH I29K53.76.17E+056.75E−041.10E−0914.815291.13A5.338VH I29W56.61.31E+062.34E−031.78E−0914.94381.13A5.340VH S3OL48.76.51E+051.47E−042.25E−1015.6671531.13A5.341VH S30Y54.37.54E+051.37E−041.82E−1017.3721031.13A5.342VH S3OD54.96.17E+051.23E−042.00E−1017.4811031.13A5.347VH N31A53.87.53E+056.75E−048.97E−1017.3151181.13A5.348VH N31S54.67.82E+055.03E−046.44E−1017.7201071.13A5.350VH N31Y56.29.01E+058.80E−049.76E−1017.611NE1.13A5.351VH N31D59.86.40E+052.03E−043.17E−1019.249751.13A5.352VH N31Q54.86.93E+054.21E−046.07E−1017.7241561.13A5.353VH N31K55.56.85E+057.54E−041.10E−0917.7131701.13A5.354VH N31H53.77.63E+051.32E−041.73E−1017.5751431.13A5.356VH G32A52.35.25E+053.02E−045.76E−1016.4331791.13A5.357VH G32S49.26.02E+051.23E−032.04E−0915.381531.13A5.495VH S94K556.76E+052.91E−044.31E−1017.6341271.13A5.497VH S94W56.81.67E+062.92E−031.75E−0912.53381.13A5.498VH S94P55.54.45E+051.17E−042.63E−101685171.13A5.500VH L95S2.7NENE1.13A5.504VH L95K51.71.53E+063.12E−032.03E−0911.33671.13A5.505VH L95H52.95.59E+056.66E−041.19E−0916.315881.13A5.507VH L95P54.7NB401.13A5.509VH G96S54.45.94E+051.92E−043.23E−101752881.13A5.510VH G96L544.11E+067.90E−031.92E−098.51631.13A5.513VH G96Q54.84.45E+055.13E−041.15E−0916.519641.13A5.514VH G96K54.95.14E+066.84E−031.33E−0911.411471.13A5.515VH G96H53.24.37E+051.20E−032.74E−0915.381171.13A5.516VH G96W531.31E+061.40E−021.07E−086.71551.13A5.517VH G96P47.81.35E+055.04E−043.73E−0911.720151.13A5.518VH N97A525.35E+051.72E−043.21E−1016.358951.13A5.520VH N97L54.54.73E+051.11E−042.35E−101689961.13A5.521VH N97Y54.75.05E+053.34E−046.62E−1016.130751.13A5.523VH N97Q53.75.06E+059.79E−051.94E−1016.61011271.13A5.524VH N97K52.52.89E+051.37E−044.75E−1015.1721671.13A5.525VH N97H53.14.11E+052.14E−045.20E−1015461131.13A5.526VH N97W53.75.28E+055.26E−049.95E−1015.519561.13A5.527VH N97P55.14.21E+054.38E−041.04E−0915.523681.13A5.528VH W98A57.67.26E+051.87E−042.57E−1018.4531141.13A5.529VH W98S53.96.23E+052.36E−043.78E−1016.842831.13A5.530VH W98L54.33.38E+051.72E−045.08E−1014.958381.13A5.531VH W98Y56.37.15E+051.12E−041.56E−1018.1881431.13A5.532VH W98D56.42.17E+058.87E−054.09E−1014.6112421.13A5.533VH W98Q53.85.01E+051.54E−043.08E−1015.864511.13A5.534VH W98K53.43.46E+058.25E−042.38E−0911.512731.13A5.535VH W98H53.86.13E+051.45E−042.37E−101768811.13A5.536VH W98P56.19.97E+048.17E−048.20E−0914.212311.13A5.537VH F99A54.21.54E+054.90E−043.18E−0913.720371.13A5.538VH F99S53.31.18E+051.02E−038.67E−099.910401.13A5.539VH F99L54.27.05E+051.15E−041.63E−1016.786551.13A5.540VH F99Y55.65.27E+059.13E−051.73E−1014.9109411.13A5.541VH F99D53.91.46E+051.11E−027.56E−084.41321.13A5.542VH F99Q53.89.93E+041.00E−031.01E−081310351.13A5.543VH F99K54.6NB381.13A5.544VH F99H54.82.05E+059.50E−054.64E−1014.5104411.13A5.545VH F99W54.55.35E+044.09E−047.66E−09624291.13A5.546VH F99P53.1NB431.13A5.547VH D101A55.61.68E+051.40E−048.37E−1014.771691.13A5.548VH D101S55.22.38E+051.11E−044.67E−1015.389781.13A5.549VH D101L53.65.94E+054.76E−048.01E−1016.321431.13A5.551VH D101Q54.55.93E+052.19E−043.69E−1017.145781.23A5.0401Wild type50.37.40E+051.33E−041.80E−1016.31.23A5.0402Wild type50.37.14E+058.22E−051.15E−1016.11.23A5.0403Wild type507.28E+051.01E−041.39E−1016.11.23A5.0404Wild type50.27.32E+051.10E−041.50E−1016.21.23A5.0405Wild type50.17.27E+059.36E−051.29E−1016.11.23A5.040AverageWild type50.27.28E+051.04E−041.43E−1016.21001001.2IgG450.8N / AN / AN / A0N / AIsotype1.23A5.040Wild type52.67.35E+051.11E−041.51E−1016.694(Purified)1.23A5.553VH D101H51.94.41E+051.63E−043.69E−1015.564661.23A5.555VH D101P51.37.54E+038.00E−041.06E−0723.613391.23A5.556VH Y102A50.66.60E+058.94E−051.35E−1016.41161061.23A5.557VH Y102S53.56.85E+051.11E−041.61E−1017.2941351.23A5.558VH Y102L53.27.40E+051.14E−041.54E−1016.891511.23A5.559VH Y102D52.87.03E+051.41E−042.01E−1016.774Si1.23A5.560VH Y102Q52.67.14E+051.08E−041.51E−1016.996871.23A5.561VH Y102K55.97.23E+058.01E−051.11E−1017.7130901.23A5.562VH Y102H50.47.22E+051.00E−041.39E−1016.41041321.23A5.563VH Y102W50.97.51E+051.16E−041.54E−1016.8901171.23A5.564VH Y102P536.44E+051.29E−042.01E−101581322.13A5.0401Wild type68.57.13E+051.16E−041.63E−1022.62.13A5.040AverageWild type68.57.13E+051.16E−041.63E−1022.61001002.1IgG460.7N / AN / AN / A1.6N / AIsotype2.13A5.040Wild type75.67.37E+051.05E−041.42E−1024.3110(Purified)2.13A5.048VL G24A697.16E+059.18E−051.28E−1022.71261042.13A5.049VL G24S72.56.98E+051.03E−041.47E−1023.71131042.13A5.050VL G24L73.86.79E+058.13E−051.20E−1023.91431032.13A5.051VL G24Y61.76.40E+051.12E−041.75E−1020.3104912.13A5.052VL G24D66.77.16E+059.71E−051.36E−1021.91191042.13A5.053VL G24Q72.66.99E+059.84E−051.41E−1023.6118952.13A5.054VL G24K74.27.12E+059.59E−051.35E−1024.1121982.13A5.055VL G24H60.96.90E+059.17E−051.33E−1020.2126992.13A5.056VL G24W64.46.41E+058.06E−051.26E−1021.11441112.13A5.057VL G25A69.97.04E+058.75E−051.24E−1022.91331092.13A5.059VL G25L63.17.16E+051.04E−041.45E−1021112832.13A5.060VL G25Y67.77.05E+058.55E−051.21E−1022.2136962.13A5.061VL G25D71.97.26E+051.11E−041.53E−1023.4105942.13A5.062VL G25Q73.26.52E+059.24E−051.42E−1023.5126942.13A5.064VL G25H60.46.90E+059.21E−051.33E−1020.11261082.13A5.065VL G25W70.87.37E+058.97E−051.22E−1023.1129812.13A5.066VL N26A76.87.27E+058.75E−051.20E−1024.7133772.13A5.069VL N26Y71.26.79E+058.73E−051.28E−1022.91331002.13A5.070VL N26D62.77.22E+057.87E−051.09E−1020.7147952.13A5.071VL N26Q67.47.05E+051.08E−041.53E−1022107952.13A5.072VL N26K69.77.04E+051.17E−041.66E−1022.799992.13A5.073VL N26H74.87.15E+051.11E−041.55E−1024105952.13A5.074VL N26W62.66.41E+059.82E−051.53E−1020.3118872.13A5.075VL N27A62.76.55E+059.29E−051.42E−1020.71251202.13A5.076VL N27S71.77.04E+051.09E−041.55E−1023.2106932.13A5.077VL N27L71.76.73E+058.98E−051.33E−1023.41291022.13A5.078VL N27Y61.56.16E+051.02E−041.65E−1020.31141032.13A5.080VL N27Q68.36.52E+051.07E−041.64E−1022.1108912.13A5.081VL N27K71.56.89E+058.84E−051.28E−1023.2131962.13A5.082VL N27H74.77.06E+057.85E−051.11E−1024.1148902.13A5.084VL I28A62.16.71E+056.20E−059.24E−1120.3187912.13A5.085VL I28S91.96.37E+051.16E−041.82E−1031.3100472.13A5.086VL I28L81.56.02E+051.04E−041.73E−1027.2112622.13A5.087VL I28Y73.17.07E+058.52E−051.20E−1023.8136932.13A5.088VL I28D80.35.56E+059.83E−051.77E−1027.9118402.13A5.089VL I28Q79.85.80E+051.03E−041.77E−1027.8113542.13A5.093VL G29A68.45.42E+051.10E−042.04E−1022.6105592.13A5.094VL G29S67.66.98E+059.31E−051.33E−1021.9119932.23A5.0401Wild type52.47.77E+051.05E−041.35E−1017.92.23A5.0402Wild type52.77.89E+051.09E−041.38E−1017.92.23A5.0403Wild type52.77.72E+051.37E−041.77E−1018.12.23A5.040AverageWild type52.67.79E+051.17E−041.50E−1017.91001002.2IgG456.6N / AN / AN / A0.5N / AIsotype2.23A5.040Wild type54.97.93E+051.06E−041.33E−1018.7110(Purified)2.23A5.095VL G29L55.47.17E+059.08E−051.27E−1018.7122752.23A5.096VL G29Y56.76.92E+051.33E−041.92E−1019.283582.23A5.097VL G29D56.27.31E+057.19E−059.84E−1118.7154852.23A5.099VL G29K49.97.18E+059.08E−051.26E−1017.2122712.23A5.100VL G29H527.38E+058.65E−051.17E−1017.71281002.23A5.101VL G29W55.87.59E+059.40E−051.24E−1018.71181142.23A5.102VL S30A56.36.85E+051.09E−041.58E−1018.7102842.23A5.103VL S3OL49.57.87E+059.94E−051.26E−1016.91121262.23A5.104VL S30Y54.78.17E+051.08E−041.33E−1018.5103932.23A5.105VL S3OD56.48.20E+058.91E−051.09E−1018.81251192.23A5.106VL S30Q58.37.22E+059.10E−051.26E−1019.7122782.23A5.107VL S3OK50.67.85E+057.34E−059.34E−1117.31511022.23A5.113VL K31Y53.88.35E+058.73E−051.05E−1018.4127992.23A5.114VL K31D54.54.89E+054.73E−049.65E−1017.9231052.23A5.115VL K31Q57.94.65E+054.60E−049.91E−101924852.23A5.116VL K31H49.55.52E+052.46E−044.46E−1016.7451042.23A5.117VL K31W52.75.34E+051.33E−042.49E−1017.7831252.23A5.118VL N32A55.85.07E+056.39E−041.26E−0918.317672.23A5.121VL N32Y56.19.57E+059.68E−051.01E−1019.1115912.23A5.122VL N32D49.85.68E+051.25E−042.20E−1015.8891152.23A5.123VL N32Q53.98.85E+051.25E−041.41E−1018.6891022.23A5.125VL N32H56.49.01E+057.73E−058.58E−1119.2144902.23A5.126VL N32W56.66.35E+059.26E−051.46E−1018.8120792.23A5.127VL V33A49.17.15E+058.05E−051.13E−1016.6138742.23A5.128VL V33S51.52.85E+059.65E−053.38E−1013.71151262.23A5.130VL V33Y56.17.41E+059.16E−051.24E−1019121682.23A5.131VL V33D52.47.40E+059.67E−051.31E−1019.3115402.23A5.132VL V33Q48.76.61E+051.38E−042.09E−1016.780502.23A5.135VL V33W48.45.26E+051.04E−041.99E−1018.8107242.23A5.137VL Y34S48.36.96E+059.88E−051.42E−1019112322.23A5.138VL Y34L49.45.94E+052.16E−043.64E−1016.651562.23A5.139VL Y34D52.84.63E+054.90E−041.06E−0913.123682.23A5.140VL Y34Q54.47.01E+055.40E−047.71E−1018.221552.23A5.141VL Y34K568.16E+053.72E−044.56E−1018.830642.23A5.142VL Y34H48.86.71E+055.00E−047.46E−1016.322822.23A5.143VL Y34W53.56.98E+052.19E−033.13E−099.75582.23A5.165VL S52K52.65.32E+059.30E−041.75E−0916.6121672.23A5.168VL D53A57.21.20E+062.04E−031.71E−0917.15512.23A5.172VL D53Q50.37.13E+051.08E−041.51E−1017.1103972.23A5.174VL D53H537.45E+051.23E−041.65E−1017.7901202.23A5.186VL P55S55.87.97E+051.08E−041.35E−1018.61031082.23A5.199VL S56K57.37.71E+051.18E−041.54E−1018.8941042.23A5.201VL S56W49.67.41E+051.24E−041.67E−1017.190922.23A5.210VL Q89P53.53.80E+054.78E−041.26E−091825722.23A5.211VL V90A55.77.65E+051.06E−041.39E−1018.8105783.13A5.0401Wild type53.37.72E+051.32E−041.71E−1018.23.13A5.0402Wild type53.47.76E+051.15E−041.48E−1018.13.13A5.0403Wild type53.37.73E+051.03E−041.34E−1018.13.13A5.0404Wild type547.69E+051.14E−041.49E−1018.53.13A5.0405Wild type53.87.84E+051.20E−041.53E−1018.43.13A5.040AverageWild type53.67.75E+051.17E−041.51E−1018.21001003.1IgG456N / AN / AN / A0.7N / AIsotype3.13A5.040Wild type52.58.07E+051.40E−041.74E−1017.983(Purified)3.13A5.213VL V9OL53.77.49E+051.55E−042.06E−1018.275583.13A5.214VL V90Y546.50E+052.92E−044.49E−101840853.13A5.215VL V9OD52.95.54E+054.19E−047.56E−1019.928273.13A5.216VL V90Q48.65.93E+051.88E−043.18E−1016.362743.13A5.217VL V9OK44.36.34E+058.30E−051.31E−1017141483.13A5.218VL V9OH47.55.90E+052.99E−045.07E−1017.639603.13A5.219VL V9OW53.45.69E+056.64E−041.17E−0917.918773.13A5.220VL V9OP47.72.72E+057.61E−032.79E−081.72763.13A5.221VL W91A51.83.05E+057.08E−032.32E−082.42793.13A5.223VL W91L53.62.59E+062.55E−029.83E−097.70983.13A5.224VL W91Y45.18.82E+059.03E−041.02E−0915.813643.13A5.226VL W91Q51.15.38E+063.79E−027.05E−096.80753.13A5.227VL W91K51.8NB713.13A5.228VL W91H50.91.53E+067.24E−034.74E−0992623.13A5.229VL W91P39.44.02E+057.40E−031.84E−081.42363.13A5.230VL D92A51.25.63E+059.94E−051.77E−1016.51181133.13A5.231VL D92S53.27.67E+059.46E−051.23E−1017.51241133.13A5.232VL D92L54.39.17E+057.59E−058.27E−11181541303.13A5.233VL D92Y47.71.11E+061.10E−049.92E−1116.4106723.13A5.234VL D92Q50.95.25E+051.39E−042.64E−1015.7841943.13A5.235VL D92K54.77.63E+058.84E−051.16E−1018132923.13A5.236VL D92H54.37.93E+051.09E−041.37E−1017.61071223.13A5.237VL D92W0.5NENE3.13A5.238VL D92P46.85.21E+051.43E−042.75E−1016.182633.13A5.239VL S93A52.23.12E+063.65E−031.17E−0915.931123.13A5.240VL S93L54.82.39E+072.45E−011.03E−086.501243.13A5.241VL S93Y50.16.20E+057.48E−031.21E−08221183.13A5.242VL S93D51.65.18E+051.68E−023.25E−082.91733.13A5.243VL S93Q54.41.90E+061.51E−027.95E−098.211113.13A5.244VL S93K55.5NB1623.13A5.245VL S93H49.72.92E+062.23E−027.64E−097.311333.13A5.246VL S93W52.83.27E+054.33E−031.33E−080.831153.13A5.247VL S93P53.86.94E+055.33E−027.68E−081.701123.13A5.248VL S94A56.18.15E+051.22E−041.50E−1019.1961013.13A5.249VL S94L49.77.81E+055.00E−046.41E−1017.123903.13A5.253VL S94K536.74E+051.48E−042.20E−1017.9791093.13A5.254VL S94H53.76.24E+051.51E−042.42E−1018.1771043.13A5.256VL S94P49.76.97E+051.27E−041.82E−1016.7921393.13A5.257VL S95A52.17.34E+051.19E−041.62E−1017.798993.13A5.258VL S95L547.73E+051.13E−041.47E−1018.3104993.13A5.259VL S95Y55.78.35E+051.07E−041.28E−1018.7109833.13A5.260VL S95D49.26.07E+051.25E−042.07E−1016.5941093.13A5.261VL S95Q48.67.87E+059.19E−051.17E−1017.8127623.13A5.262VL S95K53.17.99E+051.01E−041.26E−1018.21161183.13A5.263VL S95H55.47.83E+051.00E−041.28E−1018.71171013.13A5.264VL S95W497.95E+051.27E−041.60E−1016.592873.13A5.265VL S95P52.76.45E+055.55E−038.61E−097.121473.13A5.266VL D95aA53.75.78E+068.07E−031.40E−0910.411093.13A5.267VL D95aS55.34.53E+051.21E−032.68E−0916.7101013.13A5.268VL D95aL48.59.19E+061.35E−021.47E−098.71713.13A5.269VL D95aY51.81.77E+066.74E−033.80E−0982953.13A5.270VL D95aQ53.65.26E+067.11E−031.35E−0911.421103.13A5.271VL D95aK53.83.92E+053.83E−029.77E−0820703.13A5.272VL D95aH49.72.56E+064.90E−031.92E−0910.121883.13A5.273VL D95aW53.11.27E+061.47E−021.15E−087.11943.13A5.274VL D95aP54.61.17E+063.05E−032.61E−099.841503.13A5.276VL H95bS52.18.47E+057.33E−058.66E−1119.6160613.13A5.281VL H95bK45.57.08E+059.58E−051.35E−1016.3122643.13A5.307VH G26Y48.18.73E+051.02E−041.17E−1018.3115603.13A5.321VH G27W557.64E+055.59E−047.31E−1018.421973.13A5.335VH I29Q37.95.64E+053.90E−046.92E−1014.930373.13A5.343VH S30Q51.97.55E+051.06E−041.40E−1017.71101213.13A5.344VH S3OK53.97.41E+051.39E−041.88E−1018841563.13A5.345VH S3OH54.87.34E+051.46E−041.99E−1018.780773.13A5.349VH N31L47.17.69E+057.13E−049.27E−1016.316693.13A5.355VH N31W45.38.52E+054.80E−045.63E−1017.324593.13A5.359VH G32Y53.73.28E+064.30E−031.31E−0915.53903.13A5.360VH G32D54.21.05E+066.78E−036.44E−098.121053.13A5.361VH G32Q49.53.98E+066.22E−031.56E−0911.821333.13A5.362VH G32K52.66.07E+052.20E−033.62E−097.651713.13A5.363VH G32H55.41.81E+063.01E−031.66E−0915.341143.13A5.364VH G32W52.31.46E+071.27E−028.65E−1013.51663.13A5.365VH G33A48.76.97E+059.24E−051.33E−1016.61271163.13A5.366VH G33S538.56E+052.68E−033.14E−098.341173.13A5.367VH G33L54.28.03E+053.05E−043.79E−1017.938823.13A5.368VH G33Y55.43.97E+064.08E−031.03E−0913.431073.23A5.0401Wild 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R54L51.56.60E+051.01E−041.53E−10171181355.13A5.179VL R54Y49.26.66E+051.24E−041.86E−1016.4961385.13A5.180VL R54D49.56.79E+059.89E−051.46E−1016.61201285.13A5.181VL R54Q51.36.58E+059.55E−051.45E−10171251485.13A5.182VL R54K51.56.50E+051.17E−041.79E−10171021655.13A5.183VL R54H49.16.47E+058.44E−051.30E−1016.31411375.13A5.184VL R54W506.14E+051.22E−041.99E−1015.8987.55.13A5.185VL P55A51.96.87E+051.34E−041.95E−1016.5898.95.13A5.187VL P55L49.67.00E+059.58E−051.37E−1016.31241535.13A5.188VL P55Y49.26.61E+051.19E−041.80E−1016.41001275.13A5.189VL P55D51.16.48E+059.21E−051.42E−1016.71291555.13A5.190VL P55Q48.86.55E+051.08E−041.64E−1016.41101515.13A5.191VL P55K49.36.38E+051.05E−041.64E−1016.31131375.13A5.192VL P55H51.56.51E+051.16E−041.78E−1016.71031765.13A5.193VL P55W51.56.41E+058.80E−051.37E−1016.81351605.13A5.194VL S56A49.86.41E+051.06E−041.65E−1016.11121435.13A5.195VL S56L49.66.85E+051.06E−041.54E−1016.31121315.13A5.196VL S56Y50.26.89E+051.14E−041.66E−1016.51041235.13A5.197VL S56D50.16.87E+051.19E−041.73E−1016.51001535.13A5.198VL S56Q49.66.86E+058.85E−051.29E−1016.41341315.13A5.200VL S56H49.36.83E+051.15E−041.68E−1016.31031385.13A5.207VL Q89K49.96.84E+051.27E−041.86E−1016.4941365.13A5.209VL Q89W48.65.57E+051.27E−042.28E−1015.794725.13A5.278VL H95bY49.97.65E+057.90E−051.03E−1016.31511395.13A5.279VL H95bD49.67.01E+057.84E−051.12E−1016.51522105.13A5.280VL H95bQ50.37.47E+051.51E−042.02E−1016.6791335.13A5.284VL V96A48.27.24E+051.88E−042.60E−1015.963655.13A5.286VL V96L48.46.50E+051.94E−042.98E−1015.961795.13A5.294VL V97A48.66.27E+057.68E−051.23E−10161551335.13A5.295VL V97S48.66.12E+051.07E−041.74E−1015.91111105.13A5.296VL V97L486.71E+059.60E−051.43E−1015.9124955.13A5.297VL V97Y48.46.72E+051.07E−041.60E−1016.2111905.13A5.298VL V97D47.75.94E+051.30E−042.18E−1015.892965.13A5.300VL V97K50.15.88E+051.18E−042.01E−1015.81014.75.13A5.301VL V97H49.46.34E+058.79E−051.39E−1016.11351685.13A5.063VL G25K496.90E+057.21E−051.05E−1016.2165905.13A5.067VL N26S496.52E+051.10E−041.69E−10161081625.13A5.083VL N27W49.66.03E+051.29E−042.14E−1016.4921485.13A5.091VL I28H47.75.71E+051.30E−042.27E−1015.792755.13A5.098VL G29Q49.76.53E+051.05E−041.61E−1016.21131395.13A5.108VL S3OH49.46.95E+051.02E−041.46E−1016.21171455.13A5.109VL S3OW486.77E+051.35E−042.00E−1015.5881125.13A5.119VL N32S48.68.35E+051.08E−041.29E−1016.3110985.13A5.120VL N32L49.28.72E+051.03E−041.18E−1016.41161195.13A5.129VL V33L48.85.80E+051.42E−042.45E−1015.884995.13A5.277VL H95bL50.46.79E+051.40E−042.06E−1016.6851805.13A5.302VL V97W47.77.18E+057.59E−051.06E−1015.9157735.13A5.303VL V97P48.76.99E+051.21E−041.73E−1016.29821This table contains a summary of the calculated kinetic and titre data for each antibody supernatant tested Variant antibodies with a lower kd (slower off-rate) than parent 3A5.040 antibody have a “% 3A5.040 kd”value >100% and those with a higher kd (faster off-rate) a value <100%. Variant antibodies with a higher titre than parent 3A5.040 antibody have a “% 3A5.040 titre”value >100% and those with a lower titre, a value <100%. N / A, non-applicable. NB, No binding. NE, No expression.

[0179] TABLE 4Determination of purified 3A5.040 variant antibody IL-5 binding kinetics by BIACORE ™%RunAmino acidCaptureRmax3A5.040#Antibody IDReplicatesSubstitutionlevelka (1 / Ms)kd (Vs)KD (M)(RU)kd63A5.040 (batch 6)1Wild type48.61.44E+062.32E−041.61E−101663A5.040 (batch 6)2Wild type49.41.44E+062.14E−041.48E−1015.763A5.040 (batch 6)3Wild type49.21.36E+062.61E−041.93E−101663A5.040 (batch 6)4Wild type49.21.36E+062.46E−041.80E−101663A5.040 (batch 6)5Wild type49.31.47E+062.52E−041.71E−1016.163A5.040 (batch 6)6Wild type49.21.45E+062.13E−041.47E−101663A5.040 (batch 6)7Wild type49.21.41E+062.29E−041.63E−1016.163A5.040 (batch 6)8Wild type50.91.40E+062.45E−041.75E−1016.863A5.040 (batch 6)AverageWild type49.31.42E+062.37E−041.67E−1016.11006IgG4 iso148.8N / AN / AN / A06IgG4 iso248.9N / AN / AN / A06IgG4 iso348.8N / AN / AN / A06IgG4 IsotypeAverage49.8N / AN / AN / A0N / A63A5.040 (Purified)1Wild type49.61.43E+062.39E−041.68E−1016.163A5.040 (Purified)2Wild type49.81.48E+062.85E−041.93E−1016.263A5.040 (Purified)3Wild type49.71.45E+062.32E−041.60E−1016.163A5.040 (Purified)AverageWild type49.71.45E+062.52E−041.74E−1016.19463A5.2371VL_D92W47.91.57E+062.67E−041.70E−1015.563A5.2372VL_D92W48.71.55E+062.58E−041.66E−1015.663A5.2373VL_D92W48.51.52E+062.12E−041.40E−1015.763A5.237AverageVL_D92W48.31.55E+062.46E−041.59E−1015.69663A5.1611VL_S52L49.41.47E+062.62E−041.78E−1016.263A5.1612VL_S52L49.91.43E+062.41E−041.69E−1016.263A5.1613VL_S52L49.91.40E+062.19E−041.57E−1016.463A5.161AverageVL_S52L49.71.43E+062.41E−041.68E−1016.29963A5.1691VL_D53S48.91.23E+062.43E−041.98E−1015.663A5.1692VL_D53S49.51.26E+062.44E−041.93E−1015.963A5.1693VL_D53S49.71.23E+062.46E−041.99E−1016.163A5.169AverageVL_D53549.31.24E+062.44E−041.97E−1015.89763A5.1831VL_R54H47.91.44E+062.32E−041.61E−1015.663A5.1832VL_R54H48.21.44E+062.50E−041.74E−1015.863A5.1833VL_R54H48.41.39E+062.04E−041.47E−1015.863A5.183AverageVL_R54H48.11.42E+062.29E−041.61E−1015.710463A5.1931VL_P55W48.51.32E+062.32E−041.76E−1015.763A5.1932VL_P55W48.31.34E+062.51E−041.88E−1015.363A5.1933VL_P55W48.71.29E+062.57E−041.99E−1015.663A5.193AverageVL_P55W48.51.32E+062.47E−041.88E−1015.59663A5.1981VL_S56Q49.11.42E+062.43E−041.71E−101663A5.1982VL_S56Q491.49E+062.90E−041.95E−1015.763A5.1983VL_S56Q49.31.42E+062.38E−041.68E−1015.963A5.198AverageVL_S56Q49.11.44E+062.57E−041.78E−1015.89263A5.2781VL_H95bY48.31.55E+062.53E−041.63E−1015.963A5.2782VL_H95bY48.21.62E+062.43E−041.49E−1015.763A5.2783VL_H95bY48.51.57E+062.14E−041.36E−1015.863A5.278AverageVL_H95bY48.31.58E+062.37E−041.49E−1015.810063A5.2791VL_H95bD501.58E+062.50E−041.58E−1016.363A5.2792VL_H95bD49.71.61E+062.42E−041.50E−1016.163A5.2793VL_H95bD501.61E+062.43E−041.51E−1016.463A5.279AverageVL_H95bD49.91.60E+062.45E−041.53E−1016.29763A5.2941VL_V97A49.11.31E+062.54E−041.94E−1015.963A5.2942VL_V97A48.61.31E+062.13E−041.62E−1015.763A5.2943VL_V97A48.61.37E+062.15E−041.57E−1015.663A5.294AverageVL_V97A48.71.33E+062.27E−041.71E−1015.710463A5.3011VL_V97H48.41.51E+062.76E−041.83E−1015.863A5.3012VL_V97H48.11.58E+061.06E−046.71E−1115.663A5.3013VL_V97H47.81.60E+062.45E−041.53E−1015.463A5.301AverageVL_V97H48.11.56E+062.09E−041.34E−1015.611463A5.0631VL_G25K481.37E+062.61E−041.90E−1015.763A5.0632VL_G25K48.11.36E+062.70E−041.99E−1015.563A5.0633VL_G25K47.81.42E+062.50E−041.76E−1015.363A5.063AverageVL_G25K47.91.38E+062.60E−041.88E−1015.59163A5.3021VL_V97W48.91.65E+062.35E−041.42E−101663A5.3022VL_V97W48.91.69E+062.35E−041.39E−1015.863A5.3023VL_V97W48.51.66E+062.40E−041.45E−1015.763A5.302AverageVL_V97W48.71.67E+062.37E−041.42E−1015.810063A5.2021VL_Q89A48.91.09E+062.49E−042.29E−1015.863A5.2022VL Q89A48.71.07E+061.89E−041.77E−1015.663A5.2023VL Q89A48.81.09E+062.44E−042.25E−1015.763A5.202AverageVL_Q89A48.81.08E+062.27E−042.10E−1015.710463A5.2031VL Q89S491.28E+062.82E−042.20E−1015.963A5.2032VL Q89S48.61.28E+062.26E−041.76E−1015.663A5.2033VL Q89S48.91.27E+062.36E−041.86E−1015.763A5.203AverageVL_Q89548.81.28E+062.48E−041.94E−1015.79663A5.2041VL Q89L49.95.01E+052.20E−044.38E−1014.463A5.2042VL Q89L49.95.02E+051.88E−043.74E−1014.263A5.2043VL Q89L49.94.95E+052.11E−044.26E−1014.363A5.204AverageVL_Q89L49.94.99E+052.06E−044.13E−1014.311563A5.2051VL Q89Y48.49.87E+052.42E−042.45E−1015.563A5.2052VL Q89Y48.61.04E+062.37E−042.28E−1015.663A5.2053VL Q89Y48.51.05E+062.60E−042.47E−1015.663A5.205AverageVL_Q89Y48.51.03E+062.46E−042.40E−1015.59663A5.2081VL_Q89H519.41E+052.53E−042.68E−101663A5.2082VL_Q89H51.19.45E+052.52E−042.66E−1016.163A5.2083VL_Q89H519.54E+052.88E−043.02E−101663A5.208AverageVL_Q89H519.47E+052.64E−042.79E−10169063A5.5501VH_D101Y48.31.16E+064.40E−043.81E−1015.563A5.5502VH_D101Y48.51.17E+064.54E−043.87E−1015.763A5.5503VH_D101Y48.31.19E+064.86E−044.10E−1015.563A5.550AverageVH_D101Y48.31.17E+064.60E−043.93E−1015.55163A5.5121VH_G96D50.41.70E+078.63E−035.07E−1013.763A5.5122VH_G96D50.71.88E+079.87E−035.26E−1013.863A5.5123VH_G96D50.51.28E+061.52E−031.19E−0912.563A5.512AverageVH_G96D50.51.24E+076.67E−037.41E−1013.33.563A5.1241VL_N32K50.99.94E+052.46E−042.47E−1016.363A5.1242VL_N32K51.21.05E+062.62E−042.49E−1016.563A5.1243VL_N32K50.71.02E+062.28E−042.24E−1016.363A5.124AverageVL_N32K50.91.02E+062.45E−042.40E−1016.39773A5.040 (batch 7)1Wild type51.21.37E+062.30E−041.68E−1016.473A5.040 (batch 7)2Wild type51.61.43E+062.15E−041.50E−1016.673A5.040 (batch 7)3Wild type51.61.42E+062.84E−042.00E−1016.673A5.040 (batch 7)AverageWild type51.41.41E+062.43E−041.73E−1016.51007IgG4 Isotype148.8N / AN / AN / A07IgG4 Isotype248.9N / AN / AN / A07IgG4 Isotype348.8N / AN / AN / A07IgG4 IsotypeAverage49.8N / AN / AN / A0N / A73A5.040 (Purified)1Wild type49.61.43E+062.39E−041.68E−1016.173A5.040 (Purified)2Wild type49.81.48E+062.85E−041.93E−1016.273A5.040 (Purified)3Wild type49.71.45E+062.32E−041.60E−1016.173A5.040 (Purified)AverageWild type49.71.45E+062.52E−041.74E−1016.19473A5.0681VL_N26L52.31.29E+062.68E−042.07E−1016.873A5.0682VL_N26L531.39E+062.08E−041.50E−1017.173A5.0683VL_N26L53.11.43E+062.69E−041.88E−1017.273A5.068AverageVL_N26L52.81.37E+062.48E−041.82E−10179873A5.0791VL_N27D49.31.22E+062.29E−041.88E−1016.173A5.0792VL_N27D501.29E+062.39E−041.85E−1016.473A5.0793VL_N27D49.91.25E+062.63E−042.09E−1016.273A5.079AverageVL_N27D49.71.25E+062.44E−041.94E−1016.2100This table contains a summary of the calculated kinetic data for each purified antibody tested. Variant antibodies with a lower kd (slower off-rate) than parent 3A5.040 antibody have a “% 3A5.040 kd” value >100% and those with a higher kd (faster off-rate) a value <100%. Runs 6 and 7 were transfected separately but run in the same BIACORE ™ batch, and therefore use the same isotype and 3A5.040 (purified) controls. N / A: non-applicable.

[0180] TABLE 5Purified protein expression yield for selected3A5.040 antibody variantsAmino acid% parent 3A5.040Run #Antibody IDSubstitutionpurified yield63A5.040Parental sequence10063A5.237VL_D92W 8963A5.161VL_S52L16063A5.169VL_D53S17363A5.183VL_R54H17263A5.193VL_P55W18763A5.198VL_S56Q20563A5.278VL_H95bY21063A5.279VL_H95bD14363A5.294VL_V97A14063A5.301VL_V97H15163A5.063VL_G25K11063A5.302VL_V97W15663A5.202VL_Q89A 6463A5.203VL_Q89S12963A5.204VL_Q89L 6163A5.205VL_Q89Y 4163A5.208VL_Q89H 5463A5.550VH_D101Y 8863A5.512VH_G96D 7263A5.124VL_N32K12873A5.040Parental Sequence10073A5.068VL_N26L 9673A5.079VL_N27D 99

[0181] TABLE 6VH CDR1 Scanning MatrixVH CDR1262728293031scGGSISNA0.5 1.31 0.80.6 0.90.3 0.60.1 1.11.2 0.1S0.6 0.91 10.3 0.51.1 0.2L0.2 1.11.2 0.80.7 1.10.8 0.91.5 0.70.7 0.2Y0.6 1.10.8 0.10.5 1.20.2 0.11 0.70.6 0.1D0.6 1.20.7 0.90.7 1.10.2 01 0.80.80.5Q0.9 1.10.8 0.70.8 10.4 0.31.2 1.11.6 0.2K1 11 0.50.8 0.90.3 0.11.6 0.81.7 0.1H0.9 0.90.7 0.40.7 0.80.2 0.10.8 0.81.4 0.8W1.1 0.81 0.20.4 0.70.4 00.7 0.90.6 0.2PVH CDR13233343535A35BscGGYYWSA1.8 0.31.2 1.31.6 0.72.900.4 0.10.9 1S1.5 0.11.201.6 0.71.300.2 0.1L0.8 00.8 0.41.6 0.61.90.11 0.10.4 0.3Y0.9 01.1 01 0.50.90D1.1 02.8 01 0.20.90NE NE0.2 0.9Q1.3 0.12.8 0.11.7 0.61.300.3 0.10.5 0.2K1.7 03.5 04.1 0.23.8NB0.3 0.10.1 0H1.1 02.7 02.2 0.1200.5 0.40.8 0W0.7 01.7 0.10.5 0.52.300.5 0PSummary of difference in 3A5.040 variant antibody titre and kd for variants of VH CDR1 (AbM definition).Kabat amino acid numbering and parent 3A5.040 amino acids are given in the top row.Mutated residue identities are listed in the left column.The leftmost value in each cell is the proportional titre of each variant vs parent 3A5.040 (>1 = higher titre, <1 = lower titre), while the right value is the proportional kd (>1 = lower kd, <1 = higher kd) of each variant vs parent 3A5.040.Variants not made are blank.NB, no binding.NE, no expression. (VH CDR1 sequence disclosed as residues 26-37 of SEQ ID NO: 73).

[0182] TABLE 7VH CDR2 Scanning MatrixVH CDR2505152535455565758scYIYYSGSTYA2.70.22.40.8302.50.21.10.60.70.910.910.810S3.30.21.30.83.30.12.30.60.70.70.70.810L1.20.30.90.72.1010.21.600.70.111.20.11.20.70Y1.100.900.50.10.90.50.51.1D1.70.10.50.13.3NB1.80.2200.40.40.80.20.51.10.50Q2.40.210.22.602.10.21.50.10.40.51.11.10.610.80.1K1.40.21.20.13.5NB2.30.1200.50.51.21.30.60.910.1H30.10.902.90.32.10.510.20.40.60.410.510.90.2W1.30.11.4010.11.50.41.10.10.30.210.40.40.80.80.2PSummary of difference in 3A5.040 variant antibody titre and kd for variants of VH CDR2 (AbM definition). Kabat amino acid numbering and parent 3A5.040 amino acids are given in the top row. Mutated residue identities are listed in the left column. The leftmost value in each cell is the proportional titre of each variant vs parent 3A5.040 (>1 = higher titre, <1 = lower titre), while the right value is the proportional kd (>1 = lower kd, <1 = higher kd) of each variant vs parent 3A5.040. Variants not made are blank. NB, no binding. NE, no expression. (VH CDR2 sequence disclosed as residues 52-60 of SEQ ID NO: 73).

[0183] TABLE 8VH CDR3 Scanning MatrixVH CDR393949596979899101102scASLGNWFDYA11.20.500.80.810.61.10.50.40.20.70.71.11.2S0.61.1NE NE0.90.50.80.50.80.40.40.10.80.91.40.9L0.20.50.40.10.6010.90.40.60.60.90.40.20.50.9Y0.1NB0.30.1100.6NB0.80.31.40.90.41.10.90.5DNENENENE0.2NB0.700.90.50.41.10.300.50.7Q0.410.50.20.20.20.60.21.310.50.60.40.10.80.50.91K0.2NB1.30.30.701.501.70.70.70.10.4 NB0.70.30.91.3H0.3NB0.700.90.11.20.11.10.50.80.70.410.70.61.31W0.2NB0.40.00.700.600.60.20.30.20.30.21.20.9P0.1NB0.20.80.4NB0.20.20.70.20.30.10.4 NB0.40.10.30.8Summary of difference in 3A5.040 variant antibody titre and kd for variants of VH CDR3 (AbM definition). and Kabat amino acid positions 93 and 94. Kabat amino acid numbering and parent 3A5.040 amino acids are given in the top row. Mutated residue identities are listed in the left column. The leftmost value in each cell is the proportional titre of each variant vs parent 3A5.040 (>1 = higher titre, <1 = lower titre), while the right value is the proportional kd (>1 = lower kd, <1 = higher kd) of each variant vs parent 3A5.040. Variants not made are blank. NB, no binding. NE, no expression. (VH CDR3 sequence disclosed as residues 98-106 of SEQ ID NO: 73).

[0184] TABLE 9VL CDR1 Scanning MatrixVL CDR1242526272829scGGNNIGA1 1.31.1 1.30.8 1.31.21.20.9 1.90.6 1.1S11.10.4 1.11.6 1.10.91.10.5 10.9 1.2L1 1.40.81.11 111.30.6 1.10.8 1.3Y0.9 11 1.41 1.311.10.9 1.40.6 0.9D1 1.20.9 11 1.5110.4 1.20.9 1.6Q1 1.20.9 1.31 1.10.91.10.5 1.11.4 1.1K1 1.20.9 1.71 111.30.5 1.10.7 1.3H1 1.31.1 1.31 10.91.50.8 0.91 1.4W1.1 1.40.8 1.30.9 1.21.50.90.4 0.91.1 1.2PVL CDR13031323334scSKNVYA0.8 1.11.20.60.7 0.20.7 1.51.70.3S1.1 0.81 1.11.3 1.20.3 1.2L1.3 1.21.3 0.31.2 1.21 0.80.6 0.5Y0.9 1.11 1.30.91.20.7 1.3D1.2 1.31.1 0.21.2 0.90.4 1.20.7 0.2Q0.8 1.30.9 0.310.90.5 0.80.6 0.2K1 1.61.3 10.1 10.6 0.3H1.5 1.21 0.50.91.50.5 NB0.8 0.2W1.1 0.91.3 0.90.81.30.2 1.10.6 0.1PSummary of difference in 3A5.040 variant antibody titre and kd for variants of VL CDRI (AbM definition).Kabat amino acid numbering and parent 3A5.040 amino acids are given in the top row.Mutated residue identities are listed in the left column. The leftmost value in each cell is the proportional titre of each variant vs parent 3A5.040 (>1 = higher titre, <1 = lower titre), while the right value is the proportional kd (>1 = lower kd, <1 = higher kd) of each variant vs parent 3A5.040.Variants not made are blank.NB, no binding.NE, no expression. (VL CDR1 sequence disclosed as residues 23-33 of SEQ ID NO: 74).

[0185] TABLE 10VL CDR2 Scanning MatrixVL CDR250515253545556scDDSDRPSA0.80.60.90.80.80.90.50.11.41.20.10.91.41.1S0.910.70.71.51.91.31.11.11L0.90.40.80.61.11.81.41.31.41.21.51.21.31.1Y0.80.50.80.61.11.12.11.11.41.01.311.21D1.81.21.31.21.61.31.51Q0.70.50.90.90.11111.51.21.51.11.31.3K0.70.60.70.91.70.11.81.11.71.01.41.110.9H10.60.90.61.71.11.20.91.41.41.811.41W0.70.40.60.51.71.11.81.20.11.01.61.40.90.9PSummary of difference in 3A5.040 variant antibody titre and kd for variants of VL CDR2 (AbM definition). Kabat amino acid numbering and parent 3A5.040 amino acids are given in the top row. Mutated residue identities are listed in the left column. The leftmost value in each cell is the proportional titre of each variant vs parent 3A5.040 (>1 = higher titre, <1 = lower titre), while the right value is the proportional kd (>1 = lower kd, <1 = higher kd) of each variant vs parent 3A5.040. Variants not made are blank. NB, no binding. NE, no expression. (VL CDR2 sequence disclosed as residues 49-55 of SEQ ID NO: 74).

[0186] TABLE 11VL CDR3 Scanning MatrixVL CDR3899091929394scQVWDSSA0.610.810.801.11.21.1011S1.3100.4101.11.2L0.61.20.60.8101.31.51.200.90.2Y0.410.90.40.60.10.71.11.201.50.3D0.11.20.30.30.700.700.10.2Q0.80.60.801.90.81.101.40.6K1.40.90.51.40.7NB0.91.31.6NB1.10.8H0.50.90.60.40.601.21.11.3010.8W0.70.90.80.2NENE1.200.20.4P0.70.20.800.400.60.81.101.40.9VL CDR39595A95B9697scSDHVV111.100.20.70.7 0.61.31.510.10.61.60.2 0.51.1 1.1110.701.80.90.8 0.61 1.20.81.1101.41.50.6 0.10.91.11.10.92.11.50.1 0.11 0.90.61.31.101.30.80.3 0.20.7 1.21.21.20.700.61.20.2 00111.21.900.5 NB1.7 1.40.90.90.901.510.200.71.61.501.501.501.1 0.20.21Summary of difference in 3A5.040 variant antibody titre and kd for variants of VL CDR3 (AbM definition).Kabat amino acid numbering and parent 3A5.040 amino acids are given in the top row.Mutated residue identities are listed in the left column.The leftmost value in each cell is the proportional titre of each variant vs parent 3A5.040 (>1 = higher titre, <1 = lower titre), while the right value is the proportional kd (>1 = lower kd, <1 = higher kd) of each variant vs parent 3A5.040.Variants not made are blank.NB, no binding.NE, no expression. (VL CDR3 sequence disclosed as residues 88-98 of SEQ ID NO: 74).Expression Vector Construction

[0187] Nucleotide sequence can influence gene expression and subsequent protein expression level. Several different nucleotide sequences were examined for optimal expression of 3A5.046. These sequences are summarized in the following table:

[0188] TABLE 12Nucleotide sequenceGene NameSEQ ID Nos:3A5.046 VHSEQ ID NO: 693A5.046 VLSEQ ID NO: 713A5.046 HC constant regionSEQ ID NO: 703A5.046 LC constant regionSEQ ID NO: 72Measurement of the Affinity of Antibody 3A5.046 for Human Recombinant IL-5 by SPR

[0189] The affinity of antibody 3A5.046 for recombinant human IL-5 was determined using a BIACORE™ T200 system (GE HEALTHCARE®). Briefly, a commercial mouse anti-human IgG antibody (THERMO SCIENTIFIC®) was coupled to two adjacent (control and test) flowcells on a BIACORE™ CM5 chip (GE HEALTHCARE®) according to the manufacturer's recommendations. The 3A5.046 antibody was captured at a low density (approximately 125 RU) on the test flowcell surface and dilutions of recombinant human IL-5 or a buffer blank were injected over both test and control flowcells, at 30 μL / min (FIG. 6, Table 13).

[0190] The resulting sensorgrams were double-referenced (test flowcell values subtracted from control (anti-human IgG-coupled surface with no coated antibody) flowcell values and also a buffer blank). Subtracted sensorgrams from injections of human IL-5 at 2.5, 1.25, 0.625, 0.313, and 0.156 μg / mL were fitted to a 1:1 Langmuir binding model using BIACORE™ Evaluation software to determine binding constants from duplicate assays (Table 13).

[0191] TABLE 13Kinetic constants from a multi-concentration BIACORE ™ kineticanalysis of 3A5.046 antibody binding to recombinanthuman IL-5 at 2.5, 1.25, 0.625, 0.313 and 0.156 μg / mLAntibodyExperimentka (1 / Ms)kd (1 / s)KD (pM)Chi2 (RU2)3A5.046Repeat 18.16E+051.58E−0519.40.0269Repeat 28.49E+051.84E−0521.70.0296

[0192] These results demonstrate that antibody 3A5.046 has a high affinity for recombinant human IL-5.Potency of 3A5.046 Compared to Mepolizumab in the TF-1.6G4 Cell Line Proliferation Assay

[0193] 3A5.046 and mepolizumab were run in the TF-1.6G4 cell line proliferation assay. 3A5.046 and mepolizumab inhibited human IL-5 proliferation with a mean IC50 of 13.8 pM and 534 pM respectively (See Table 14 and FIG. 7A). Both antibodies demonstrated inhibition of cynomolgus monkey IL-5 on the TF1.6G4 cell line (FIG. 7B), with a mean IC50 for 3A5.046 of 43.8 pM and for mepolizumab of 584 pM.

[0194] TABLE 14Number ofMeanStd.AntibodyAgonistObservations(pM)Dev.MepolizumabCynomolgus4584.0982129.9019IL-5Human IL-54534.2803150.25463A5.046Cynomolgus8 43.80916 12.62095IL-5Human IL-58 13.83667  4.219783Detection of Native IL5 from Primary Cells Via ELISA Using Anti-IL-5 Antibodies of the Invention

[0195] Antibody 3A5 was used as a capture reagent in a sandwich ELISA. Paired with a commercially available detection antibody (TRFk5R; R&D SYSTEMS®, #MAB405) quantification of recombinant IL-5 levels in spiked buffer samples could be determined (FIG. 8). This method was then applied to the detection of native IL-5 secreted by primary cells. CD4+ T-cells from four donors were stimulated to secrete IL-5. The IL-5 levels were then determined using the sandwich ELISA with antibody 3A5 as the capture reagent. Three out of four donor T-cells responded to T-cell stimulation and had detectable IL-5 levels in the cell culture supernatant (FIG. 8). Similar studies were performed on human IL-5 secreted from primary human donor CD4+ T-cells using antibody 3A5.046 as a capture agent, and the results obtained were consistent with the results obtained with antibody 3A5 (data not shown).Anti-IL-5 Antibody Inhibition of Eosinophil Differentiation

[0196] A survival assay using CD34+ cord blood cells was used to assess the capacity of anti-IL-5 antibodies to inhibit the IL-5 dependent differentiation of bone marrow progenitors to eosinophils and to assess the capacity of anti-IL-5 antibodies to inhibit phenotypically mature eosinophil survival. In IL-5 treated control cells, phenotypically mature eosinophils were the predominant population at day 28 (FIG. 9, shaded region in graph). Antibody 3A5.046 was able to inhibit the differentiation of CD34+ cord blood cells, incubated with IL-5 for 28-days, into eosinophils. Antibody 3A5.046 was a more potent inhibitor of IL-5 mediated eosinophil differentiation than mepolizumab (FIG. 9).X-Ray Crystallography Analysis

[0197] The molecular structure of the complex between antibody 3A5.046 Fab and recombinant human IL-5 is investigated by X-ray crystallography. To prepare for X-ray crystallography experiments, 3A5.046 is digested by papain, which cleaves the intact antibody in the hinge region, separating the Fc and Fab domains. The 3A5.046 Fab is purified by standard Protein A affinity chromatography and complexed with recombinant non-glycosylated human IL-5. The 3A5.046 Fab:human IL-5 complex is purified for crystallization experiments using standard size-exclusion chromatography. The complex is set up in variable concentrations, into commercially available sparse-matrix crystallization screens for broad sampling of crystallization conditions. Optimisations of any ‘hit’ conditions that provide crystals of the complex from the sparse-matrix screens are performed to improve upon crystal morphology and crystal diffraction as required.

[0198] Crystals of the 3A5.046 Fab:human IL-5 complex are harvested, flash-frozen in liquid nitrogen and transported to a synchrotron facility for diffraction tests and data collection. Crystal diffraction testing is performed at a synchrotron, which houses high energy X-ray micro beam lines for producing high-resolution diffraction. In this way, X-ray diffraction data on crystals of 3A5.046 Fab:human IL-5 complex is collected. An initial structure model of the complex may be obtained using standard molecular replacement techniques with published molecular structures of IL-5 and Fab as templates. The final structure of the 3A5.046 Fab and human IL-5 complex may be obtained by iterative refinement of the structure model, until the errors of fitting are better than 30% (final Rfree of 0.3) and chemical bonding parameters (e.g. the distribution of bond lengths and angles, stereochemistry and hydrogen bonds) are within acceptable limits.Characterisation of Antibody 3A5.046 Binding to IL-5 from Non-Human Model Species

[0199] Antibody 3A5.046 was tested by BIACORE™ assay using a T200 system (GE HEALTHCARE®) for binding to recombinant cytokines human IL-5, rat IL-5, mouse IL-5, and guinea pig IL-5. This panel of non-human IL-5s was chosen to represent major animal model species used in preclinical development. An isotype control antibody (IgG4) and antibody 3A5.046 were immobilized on flowcells 1 and 2 of a Series S CM5 chip (GE HEALTHCARE®), respectively, using standard amine coupling methods. Cytokines were diluted to 1 and 10 μg / ml in HBS-EP+ running buffer (GE HEALTHCARE®) and injected across flowcells 1 and 2. Buffer-only injections across flowcells 1 and 2 were included for double-referencing. Data analysis was performed using BIACORE™ T200 evaluation software. The resulting sensorgrams were double-referenced (test flowcell values subtracted from control flowcell values and also a buffer blank).

[0200] Antibody 3A5.046 was found to bind to human IL-5 and cynomolgus monkey IL-5, but not to mouse IL-5, rat IL-5, or guinea pig IL-5 (FIG. 10A-FIG. 10E). This indicated that 3A5.046 could be used in cynomolgus macaque preclinical models.Antibody 3A5.046 does not Bind to Other Human Cytokines Closely Related to IL-5

[0201] Antibody 3A5.046 was tested by BIACORE™ assay using a T200 system (GE HEALTHCARE®) for binding to recombinant human cytokines IL-2, IL-3, IL-4, IL-5, granulocyte macrophage colony stimulating factor (GM-CSF), macrophage colony stimulating factor (M-CSF), and growth hormone (GH). This representative panel of closely-related human cytokines was chosen based on their structural and functional similarity to human IL-5. An isotype control antibody (IgG4) and antibody 3A5.046 were immobilized on flowcells 1 and 2 of a Series S CM5 chip (GE HEALTHCARE®), respectively, using standard amine coupling methods. Cytokines were diluted to 1 and 10 μg / ml in HBS-EP+ running buffer (GE HEALTHCARE®) and injected across flowcells 1 and 2. Buffer-only injections across flowcells 1 and 2 were included for double-referencing. Data analysis was performed using BIACORE™ T200 evaluation software. The resulting sensorgrams were double-referenced (test flowcell values subtracted from control flowcell values and also a buffer blank).

[0202] Antibody 3A5.046 was found to bind only to human IL-5 and not human cytokines IL-2, IL-3, IL-4, GM-CSF, M-CSF, or GH (FIG. 11A-FIG. 11D), thus demonstrating high specificity of 3A5.046 for human IL-5.Evaluation of Antibody 3A5.046 in an Ascaris suum-Induced Cynomolgus Monkey Airways Eosinophilia Model

[0203] Antibody 3A5.046 was tested using the Ascaris suum cynomolgus model of acute airways eosinophilia as described in Hart, T. K., et al., Preclinical efficacy and safety of mepolizumab (SB-240563), a humanized monoclonal antibody to IL-5, in cynomolgus monkeys. J Allergy Clin Immunol, 2001. 108(2): p. 250-7.

[0204] Sixteen female cynomolgus macaques that had been previously sensitized to Ascaris suum were assessed for level of retained sensitivity by intradermal challenge with A. suum. Fourteen of these animals were selected for participation in the study based on the skin wheal response to challenge and randomized into two groups of n=7 per group. Two weeks after intradermal skin wheal testing, baseline blood and bronchoalveolar lavage fluid (BALF) samples were collected (Day 8). About 24 hours after baseline sample collection, animals were treated with either placebo (comprising aqueous buffer pH 6.3 with 200 mM arginine and 25 mM histidine) or antibody 3A5.046 (10 mg / kg intravenously, in aqueous buffer pH 6.3 with 200 mM arginine and 25 mM histidine) in a blinded manner (Day 7). One week after treatment with placebo or antibody a second intradermal A. suum challenge was performed and an A. suum inhalation challenge was given to all animals by delivery of an equal amount of A. suum extract (Greer Laboratories Inc.; 5 mg / ml in sterile water) administered via PARI® LC nebulizer (Day 0). Skin wheal diameter was measured 15 minutes after challenge. Two days afterA. suum inhalation challenge, blood and BALF samples were collected. Additional blood samples for haematology endpoints and PK analysis were collected weekly starting on Day 10 until Day 45.

[0205] There was no significant difference in the skin wheal responses between the first and second intradermal A. suum challenges. All animals showed an increase in eosinophils in the BALF as a result of A. suum challenge but in animals treated with antibody 3A5.046, this increase was negligible. There were significantly fewer eosinophils in the BALF of animals treated with antibody 3A5.046, compared to placebo-treated animals 48 hours after A. suum challenge (p<0.01; Mann-Whitney test; FIG. 12).

[0206] A. suum challenge also led to significantly increased blood eosinophils over baseline in placebo-treated animals to day 10 post-challenge (FIG. 13A). Treatment with antibody 3A5.046 led to a significant decrease in blood eosinophils to levels below baseline (FIG. 13A), and this depression in blood eosinophil numbers was maintained over a period of at least 52 days after initial dose (FIG. 13B).

[0207] In summary, treatment with 3A5.046 significantly reduced A. suum-induced airways eosinophilia 48 hours after challenge, and significantly decreased blood eosinophils, to below baseline levels, for at least 52 days following a single intravenous (IV) 10 mg / kg dose.Investigation of Antibody Function in a Humanized IL-5 Rat Model of Airways Eosinophilia

[0208] Alternaria alternata (Alternaria) is a fungal allergen that is known to trigger asthma in humans and can cause eosinophilia and asthma-like symptoms in rodents. When challenged with Alternaria, humanized IL-5 knock-in rats (animals in which the rat IL-5 has been replaced by human IL-5) show elevated eosinophil levels in bronchoalveolar lavage fluid (BALF) from lungs (FIG. 14).

[0209] A single intratracheal instillation of Alternaria suspension is administered to humanized IL-5 rats, and blood and BALF samples are collected 2, 3 or 4 days after Alternaria administration. Typically in this model eosinophils are significantly increased in BALF samples and may be either increased or unchanged in blood samples. Test antibodies are administered prior to induction of eosinophilia with Altemaria and assessed for their ability to reduce eosinophil numbers in BALF and blood.

[0210] Those skilled in the art will appreciate that numerous changes and modifications can be made to the preferred embodiments of the invention and that such changes and modifications can be made without departing from the spirit of the invention. It is, therefore, intended that the appended claims cover all such equivalent variations as fall within the true spirit and scope of the invention.

[0211] The disclosures of each patent, patent application, and publication cited or described in this document are hereby incorporated herein by reference, in its entirety.

[0212] TABLE 15SequencesProtein chainSequenceProtein ChainSequenceConsensus LCDR1GX1X2X3X4X5X6KX7X8YSEQ ID NO: 1Consensus LCDR2DDX8X9RPSSEQ ID NO: 2Consensus LCDR3QVWX10SSSDX11VX12SEQ ID NO: 3Antibody 3A53A5 HCDR1GGSISNGGYYWS3A5 LCDR1GGNNIGSKNVYSEQ ID NO: 4SEQ ID NO: 53A5 HCDR2YIYYSGSTY3A5 LCDR2DDSDRPSSEQ ID NO: 6SEQ ID NO: 73A5 HCDR3LGNWFDY3A5 LCDR3QVWYSSSDHVVSEQ ID NO: 8SEQ ID NO: 93A5 VHQVQLQESGPGLVKPSQTLSLTCTVSGGSISNGGYYWSWIRQHPGKGLEWIGYIYYSGSTYYNPSLKSRVSISVDTSKNQFSLKLNSVTAADTAVYYCASLGNWFDYWGQGTLVTVSSSEQ ID NO: 103A5 VLSSILTQPPSVSVAPGQTARITCGGNNIGSKNVYWYQQKPGQAPVLVVHDDSDRPSGIPERFSGSNSGNTATLTISRVEVGDEADYSCQVWYSSSDHVVFGGGTKLTVLGSEQ ID NO: 113A5 HCQVQLQESGPGLVKPSQTLSLTCTVSGGSISNGGYYWSWIRQHPGKGLEWIGYIYYSGSTYYNPSLKSRVSISVDTSKNQFSLKLNSVTAADTAVYYCASLGNWFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGSEQ ID NO: 123A5 LCSSILTQPPSVSVAPGQTARITCGGNNIGSKNVYWYQQKPGQAPVLVVHDDSDRPSGIPERFSGSNSGNTATLTISRVEVGDEADYSCQVWYSSSDHVVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECSSEQ ID NO: 13Antibody 3A5.0013A5.001 HCDR1GGSISNGGYYWS3A5.001 LCDR1GGNNIGSKNVYSEQ ID NO: 4SEQ ID NO: 53A5.001 HCDR2YIYYSGSTY3A5.001 LCDR2DDSDRPSSEQ ID NO: 6SEQ ID NO: 73A5.001 HCDR3LGNWFDY3A5.001 LCDR3QVWYSSSDHVVSEQ ID NO: 8SEQ ID NO: 93A5.001 VHQVQLQESGPGLVKPSQTLSLTCTVSGGSISNGGYYWSWIRQHPGKGLEWIGYIYYSGSTYYNPSLKSRVSISVDTSKNQFSLKLNSVTAADTAVYYCASLGNWFDYWGQGTLVTVSSSEQ ID NO: 103A5.001 VLSSILTQPPSVSVAPGQTARITCGGNNIGSKNVYWYQQKPGQAPVLVVHDDSDRPSGIPERFSGSNSGNTATLTISRVEVGDEADYSCQVWYSSSDHVVFGGGTKLTVLGSEQ ID NO: 113A5.001 HCQVQLQESGPGLVKPSQTLSLTCTVSGGSISNGGYYWSWIRQHPGKGLEWIGYIYYSGSTYYNPSLKSRVSISVDTSKNQFSLKLNSVTAADTAVYYCASLGNWFDYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGSEQ ID NO: 143A5.001 LCSSILTQPPSVSVAPGQTARITCGGNNIGSKNVYWYQQKPGQAPVLVVHDDSDRPSGIPERFSGSNSGNTATLTISRVEVGDEADYSCQVWYSSSDHVVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECSSEQ ID NO: 13Antibody 3A5.0403A5.040 HCDR1GGSISNGGYYWS3A5.040 LCDR1GGNNIGSKNVYSEQ ID NO: 4SEQ ID NO: 53A5.040 HCDR2YIYYSGSTY3A5.040 LCDR2DDSDRPSSEQ ID NO: 6SEQ ID NO: 73A5.040 HCDR3LGNWFDY3A5.040 LCDR3QVWDSSSDHVVSEQ ID NO: 8SEQ ID NO: 153A5.040 VHQVQLQESGPGLVKPSQTLSLTCTVSGGSISNGGYYWSWIRQHPGKGLEWIGYIYYSGSTYYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCASLGNWFDYWGQGTLVTVSSSEQ ID NO: 163A5.040 VLSYVLTQPPSVSVAPGQTARITCGGNNIGSKNVYWYQQKPGQAPVLVVHDDSDRPSGIPERFSGSNSGNTATLTISRVEVGDEADYSCQVWDSSSDHVVFGGGTKLTVLGSEQ ID NO: 173A5.040 HCQVQLQESGPGLVKPSQTLSLTCTVSGGSISNGGYYWSWIRQHPGKGLEWIGYIYYSGSTYYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCASLGNWFDYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGSEQ ID NO: 183A5.040 LCSYVLTQPPSVSVAPGQTARITCGGNNIGSKNVYWYQQKPGQAPVLVVHDDSDRPSGIPERFSGSNSGNTATLTISRVEVGDEADYSCQVWDSSSDHVVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECSSEQ ID NO: 19Antibody 3A5.0463A5.046 HCDR1GGSISNGGYYWS3A5.046 LCDR1GGNNIGSKNVYSEQ ID NO: 4SEQ ID NO: 53A5.046 HCDR2YIYYSGSTY3A5.046 LCDR2DDSDRPSSEQ ID NO: 6SEQ ID NO: 73A5.046 HCDR3LGNWFDY3A5.046 LCDR3QVWDSSSDHVVSEQ ID NO: 8SEQ ID NO: 153A5.046 VHQVQLQESGPGLVKPSQTLSLTCTVSGGSISNGGYYWSWIRQHPGKGLEWIGYIYYSGSTYYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCASLGNWFDYWGQGTLVTVSSSEQ ID NO: 163A5.046 VLSYVLTQPPSVSVAPGQTARITCGGNNIGSKNVYWYQQKPGQAPVLVVHDDSDRPSGIPERFSGSNSGNTATLTISRVEVGDEADYSCQVWDSSSDHVVFGGGTKLTVLGSEQ ID NO: 173A5.046 HCQVQLQESGPGLVKPSQTLSLTCTVSGGSISNGGYYWSWIRQHPGKGLEWIGYIYYSGSTYYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCASLGNWFDYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGSEQ ID NO: 203A5.046 LCSYVLTQPPSVSVAPGQTARITCGGNNIGSKNVYWYQQKPGQAPVLVVHDDSDRPSGIPERFSGSNSGNTATLTISRVEVGDEADYSCQVWDSSSDHVVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECSSEQ ID NO: 19Antibody 3A5.0633A5.063 HCDR1GGSISNGGYYWS3A5.063 LCDR1GKNNIGSKNVYSEQ ID NO: 4SEQ ID NO: 213A5.063 HCDR2YIYYSGSTY3A5.063 LCDR2DDSDRPSSEQ ID NO: 6SEQ ID NO: 73A5.063 HCDR3LGNWFDY3A5.063 LCDR3QVWDSSSDHVVSEQ ID NO: 8SEQ ID NO: 153A5.063 VHQVQLQESGPGLVKPSQTLSLTCTVSGGSISNGGYYWSWIRQHPGKGLEWIGYIYYSGSTYYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCASLGNWFDYWGQGTLVTVSSSEQ ID NO: 163A5.063 VLSYVLTQPPSVSVAPGQTARITCGKNNIGSKNVYWYQQKPGQAPVLVVHDG25KDSDRPSGIPERFSGSNSGNTATLTISRVEVGDEADYSCQVWDSSSDHVVFGGGTKLTVLGSEQ ID NO: 223A5.063 HCQVQLQESGPGLVKPSQTLSLTCTVSGGSISNGGYYWSWIRQHPGKGLEWIGYIYYSGSTYYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCASLGNWFDYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGSEQ ID NO: 183A5.063 LCSYVLTQPPSVSVAPGQTARITCGKNNIGSKNVYWYQQKPGQAPVLVVHDG25KDSDRPSGIPERFSGSNSGNTATLTISRVEVGDEADYSCQVWDSSSDHVVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECSSEQ ID NO: 23Antibody 3A5.0703A5.070 HCDR1GGSISNGGYYWS3A5.070 LCDR1GGDNIGSKNVYSEQ ID NO: 4SEQ ID NO: 243A5.070 HCDR2YIYYSGSTY3A5.070 LCDR2DDSDRPSSEQ ID NO: 6SEQ ID NO: 73A5.070 HCDR3LGNWFDY3A5.070 LCDR3QVWDSSSDHVVSEQ ID NO: 8SEQ ID NO: 153A5.070 VHQVQLQESGPGLVKPSQTLSLTCTVSGGSISNGGYYWSWIRQHPGKGLEWIGYIYYSGSTYYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCASLGNWFDYWGQGTLVTVSSSEQ ID NO: 163A5.070 VLSYVLTQPPSVSVAPGQTARITCGGDNIGSKNVYWYQQKPGQAPVLVVHDN26DDSDRPSGIPERFSGSNSGNTATLTISRVEVGDEADYSCQVWDSSSDHVVFGGGTKLTVLGSEQ ID NO: 253A5.070 HCQVQLQESGPGLVKPSQTLSLTCTVSGGSISNGGYYWSWIRQHPGKGLEWIGYIYYSGSTYYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCASLGNWFDYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGSEQ ID NO: 183A5.070 LCSYVLTQPPSVSVAPGQTARITCGGDNIGSKNVYWYQQKPGQAPVLVVHDN26DDSDRPSGIPERFSGSNSGNTATLTISRVEVGDEADYSCQVWDSSSDHVVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECSSEQ ID NO: 26Antibody 3A5.0823A5.082 HCDR1GGSISNGGYYWS3A5.082 LCDR1GGNHIGSKNVYSEQ ID NO: 4SEQ ID NO: 273A5.082 HCDR2YIYYSGSTY3A5.082 LCDR2DDSDRPSSEQ ID NO: 6SEQ ID NO: 73A5.082 HCDR3LGNWFDY3A5.082 LCDR3QVWDSSSDHVVSEQ ID NO: 8SEQ ID NO: 153A5.082 VHQVQLQESGPGLVKPSQTLSLTCTVSGGSISNGGYYWSWIRQHPGKGLEWIGYIYYSGSTYYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCASLGNWFDYWGQGTLVTVSSSEQ ID NO: 163A5.082 VLSYVLTQPPSVSVAPGQTARITCGGNHIGSKNVYWYQQKPGQAPVLVVHDN27HDSDRPSGIPERFSGSNSGNTATLTISRVEVGDEADYSCQVWDSSSDHVVFGGGTKLTVLGSEQ ID NO: 283A5.082 HCQVQLQESGPGLVKPSQTLSLTCTVSGGSISNGGYYWSWIRQHPGKGLEWIGYIYYSGSTYYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCASLGNWFDYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGSEQ ID NO: 183A5.082 LCSYVLTQPPSVSVAPGQTARITCGGNHIGSKNVYWYQQKPGQAPVLVVHDN27HDSDRPSGIPERFSGSNSGNTATLTISRVEVGDEADYSCQVWDSSSDHVVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECSSEQ ID NO: 29Antibody 3A5.0843A5.084 HCDR1GGSISNGGYYWS3A5.084 LCDR1GGNNAGSKNVYSEQ ID NO: 4SEQ ID NO: 303A5.084 HCDR2YIYYSGSTY3A5.084 LCDR2DDSDRPSSEQ ID NO: 6SEQ ID NO: 73A5.084 HCDR3LGNWFDY3A5.084 LCDR3QVWDSSSDHVVSEQ ID NO: 8SEQ ID NO: 153A5.084 VHQVQLQESGPGLVKPSQTLSLTCTVSGGSISNGGYYWSWIRQHPGKGLEWIGYIYYSGSTYYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCASLGNWFDYWGQGTLVTVSSSEQ ID NO: 163A5.084 VLSYVLTQPPSVSVAPGQTARITCGGNNAGSKNVYWYQQKPGQAPVLVVHI28ADDSDRPSGIPERFSGSNSGNTATLTISRVEVGDEADYSCQVWDSSSDHVVFGGGTKLTVLGSEQ ID NO: 313A5.084 HCQVQLQESGPGLVKPSQTLSLTCTVSGGSISNGGYYWSWIRQHPGKGLEWIGYIYYSGSTYYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCASLGNWFDYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGSEQ ID NO: 183A5.084 LCSYVLTQPPSVSVAPGQTARITCGGNNAGSKNVYWYQQKPGQAPVLVVHI28ADDSDRPSGIPERFSGSNSGNTATLTISRVEVGDEADYSCQVWDSSSDHVVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECSSEQ ID NO: 32Antibody 3A5.1073A5.107 HCDR1GGSISNGGYYWS3A5.107 LCDR1GGNNIGKKNVYSEQ ID NO: 4SEQ ID NO: 333A5.107 HCDR2YIYYSGSTY3A5.107 LCDR2DDSDRPSSEQ ID NO: 6SEQ ID NO: 73A5.107 HCDR3LGNWFDY3A5.107 LCDR3QVWDSSSDHVVSEQ ID NO: 8SEQ ID NO: 153A5.107 VHQVQLQESGPGLVKPSQTLSLTCTVSGGSISNGGYYWSWIRQHPGKGLEWIGYIYYSGSTYYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCASLGNWFDYWGQGTLVTVSSSEQ ID NO: 163A5.107 VLSYVLTQPPSVSVAPGQTARITCGGNNIGKKNVYWYQQKPGQAPVLVVHDS30KDSDRPSGIPERFSGSNSGNTATLTISRVEVGDEADYSCQVWDSSSDHVVFGGGTKLTVLGSEQ ID NO: 343A5.107 HCQVQLQESGPGLVKPSQTLSLTCTVSGGSISNGGYYWSWIRQHPGKGLEWIGYIYYSGSTYYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCASLGNWFDYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGSEQ ID NO: 183A5.107 LCSYVLTQPPSVSVAPGQTARITCGGNNIGKKNVYWYQQKPGQAPVLVVHDS30KDSDRPSGIPERFSGSNSGNTATLTISRVEVGDEADYSCQVWDSSSDHVVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECSSEQ ID NO: 35Antibody 3A5.1253A5.125 HCDR1GGSISNGGYYWS3A5.125 LCDR1GGNNIGSKHVYSEQ ID NO: 4SEQ ID NO: 363A5.125 HCDR2YIYYSGSTY3A5.125 LCDR2DDSDRPSSEQ ID NO: 6SEQ ID NO: 73A5.125 HCDR3LGNWFDY3A5.125 LCDR3QVWDSSSDHVVSEQ ID NO: 8SEQ ID NO: 153A5.125 VHQVQLQESGPGLVKPSQTLSLTCTVSGGSISNGGYYWSWIRQHPGKGLEWIGYIYYSGSTYYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCASLGNWFDYWGQGTLVTVSSSEQ ID NO: 163A5.125 VLSYVLTQPPSVSVAPGQTARITCGGNNIGSKHVYWYQQKPGQAPVLVVHDN32HDSDRPSGIPERFSGSNSGNTATLTISRVEVGDEADYSCQVWDSSSDHVVFGGGTKLTVLGSEQ ID NO: 373A5.125 HCQVQLQESGPGLVKPSQTLSLTCTVSGGSISNGGYYWSWIRQHPGKGLEWIGYIYYSGSTYYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCASLGNWFDYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGSEQ ID NO: 183A5.125 LCSYVLTQPPSVSVAPGQTARITCGGNNIGSKHVYWYQQKPGQAPVLVVHDN32HDSDRPSGIPERFSGSNSGNTATLTISRVEVGDEADYSCQVWDSSSDHVVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECSSEQ ID NO: 38Antibody 3A5.1273A5.127 HCDR1GGSISNGGYYWS3A5.127 LCDR1GGNNIGSKNAYSEQ ID NO: 4SEQ ID NO: 393A5.127 HCDR2YIYYSGSTY3A5.127 LCDR2DDSDRPSSEQ ID NO: 6SEQ ID NO: 73A5.127 HCDR3LGNWFDY3A5.127 LCDR3QVWDSSSDHVVSEQ ID NO: 8SEQ ID NO: 153A5.127 VHQVQLQESGPGLVKPSQTLSLTCTVSGGSISNGGYYWSWIRQHPGKGLEWIGYIYYSGSTYYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCASLGNWFDYWGQGTLVTVSSSEQ ID NO: 163A5.127 VLSYVLTQPPSVSVAPGQTARITCGGNNIGSKNAYWYQQKPGQAPVLVVHDV33ADSDRPSGIPERFSGSNSGNTATLTISRVEVGDEADYSCQVWDSSSDHVVFGGGTKLTVLGSEQ ID NO: 403A5.127 HCQVQLQESGPGLVKPSQTLSLTCTVSGGSISNGGYYWSWIRQHPGKGLEWIGYIYYSGSTYYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCASLGNWFDYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGSEQ ID NO: 183A5.127 LCSYVLTQPPSVSVAPGQTARITCGGNNIGSKNAYWYQQKPGQAPVLVVHDV33ADSDRPSGIPERFSGSNSGNTATLTISRVEVGDEADYSCQVWDSSSDHVVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECSSEQ ID NO: 41Antibody 3A5.1613A5.161 HCDR1GGSISNGGYYWS3A5.161 LCDR1GGNNIGSKNVYSEQ ID NO: 4SEQ ID NO: 53A5.161 HCDR2YIYYSGSTY3A5.161 LCDR2DDLDRPSSEQ ID NO: 6SEQ ID NO: 423A5.161 HCDR3LGNWFDY3A5.161 LCDR3QVWDSSSDHVVSEQ ID NO: 8SEQ ID NO: 153A5.161 VHQVQLQESGPGLVKPSQTLSLTCTVSGGSISNGGYYWSWIRQHPGKGLEWIGYIYYSGSTYYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCASLGNWFDYWGQGTLVTVSSSEQ ID NO: 163A5.161 VLSYVLTQPPSVSVAPGQTARITCGGNNIGSKNVYWYQQKPGQAPVLVVHDS52LDLDRPSGIPERFSGSNSGNTATLTISRVEVGDEADYSCQVWDSSSDHVVFGGGTKLTVLGSEQ ID NO: 433A5.161 HCQVQLQESGPGLVKPSQTLSLTCTVSGGSISNGGYYWSWIRQHPGKGLEWIGYIYYSGSTYYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCASLGNWFDYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGSEQ ID NO: 183A5.161 LCSYVLTQPPSVSVAPGQTARITCGGNNIGSKNVYWYQQKPGQAPVLVVHDS52LDLDRPSGIPERFSGSNSGNTATLTISRVEVGDEADYSCQVWDSSSDHVVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECSSEQ ID NO: 44Antibody 3A5.1693A5.169 HCDR1GGSISNGGYYWS3A5.169 LCDR1GGNNIGSKNVYSEQ ID NO: 4SEQ ID NO: 53A5.169 HCDR2YIYYSGSTY3A5.169 LCDR2DDSSRPSSEQ ID NO: 6SEQ ID NO: 453A5.169 HCDR3LGNWFDY3A5.169 LCDR3QVWDSSSDHVVSEQ ID NO: 8SEQ ID NO: 153A5.169 VHQVQLQESGPGLVKPSQTLSLTCTVSGGSISNGGYYWSWIRQHPGKGLEWIGYIYYSGSTYYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCASLGNWFDYWGQGTLVTVSSSEQ ID NO: 163A5.169 VLSYVLTQPPSVSVAPGQTARITCGGNNIGSKNVYWYQQKPGQAPVLVVHDD53SDSSRPSGIPERFSGSNSGNTATLTISRVEVGDEADYSCQVWDSSSDHVVFGGGTKLTVLGSEQ ID NO: 463A5.169 HCQVQLQESGPGLVKPSQTLSLTCTVSGGSISNGGYYWSWIRQHPGKGLEWIGYIYYSGSTYYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCASLGNWFDYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGSEQ ID NO: 183A5.169 LCSYVLTQPPSVSVAPGQTARITCGGNNIGSKNVYWYQQKPGQAPVLVVHDD53SDSSRPSGIPERFSGSNSGNTATLTISRVEVGDEADYSCQVWDSSSDHVVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECSSEQ ID NO: 47Antibody 3A5.2323A5.232 HCDR1GGSISNGGYYWS3A5.232 LCDR1GGNNIGSKNVYSEQ ID NO: 4SEQ ID NO: 53A5.232 HCDR2YIYYSGSTY3A5.232 LCDR2DDSDRPSSEQ ID NO: 6SEQ ID NO: 73A5.232 HCDR3LGNWFDY3A5.232 LCDR3QVWLSSSDHVVSEQ ID NO: 8SEQ ID NO: 483A5.232 VHQVQLQESGPGLVKPSQTLSLTCTVSGGSISNGGYYWSWIRQHPGKGLEWIGYIYYSGSTYYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCASLGNWFDYWGQGTLVTVSSSEQ ID NO: 163A5.232 VLSYVLTQPPSVSVAPGQTARITCGGNNIGSKNVYWYQQKPGQAPVLVVHDD92LDSDRPSGIPERFSGSNSGNTATLTISRVEVGDEADYSCQVWLSSSDHVVFGGGTKLTVLGSEQ ID NO: 493A5.232 HCQVQLQESGPGLVKPSQTLSLTCTVSGGSISNGGYYWSWIRQHPGKGLEWIGYIYYSGSTYYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCASLGNWFDYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGSEQ ID NO: 183A5.232 LCSYVLTQPPSVSVAPGQTARITCGGNNIGSKNVYWYQQKPGQAPVLVVHDD92LDSDRPSGIPERFSGSNSGNTATLTISRVEVGDEADYSCQVWLSSSDHVVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECSSEQ ID NO: 50Antibody 3A5.2763A5.276 HCDR1GGSISNGGYYWS3A5.276 LCDR1GGNNIGSKNVYSEQ ID NO: 4SEQ ID NO: 53A5.276 HCDR2YIYYSGSTY3A5.276 LCDR2DDSDRPSSEQ ID NO: 6SEQ ID NO: 73A5.276 HCDR3LGNWFDY3A5.276 LCDR3QVWDSSSDSVVSEQ ID NO: 8SEQ ID NO: 513A5.276 VHQVQLQESGPGLVKPSQTLSLTCTVSGGSISNGGYYWSWIRQHPGKGLEWIGYIYYSGSTYYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCASLGNWFDYWGQGTLVTVSSSEQ ID NO: 163A5.276 VLSYVLTQPPSVSVAPGQTARITCGGNNIGSKNVYWYQQKPGQAPVLVVHDH95bSDSDRPSGIPERFSGSNSGNTATLTISRVEVGDEADYSCQVWDSSSDSVVFGGGTKLTVLGSEQ ID NO: 523A5.276 HCQVQLQESGPGLVKPSQTLSLTCTVSGGSISNGGYYWSWIRQHPGKGLEWIGYIYYSGSTYYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCASLGNWFDYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGSEQ ID NO: 183A5.276 LCSYVLTQPPSVSVAPGQTARITCGGNNIGSKNVYWYQQKPGQAPVLVVHDH95bSDSDRPSGIPERFSGSNSGNTATLTISRVEVGDEADYSCQVWDSSSDSVVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECSSEQ ID NO: 53Antibody 3A5.2783A5.278 HCDR1GGSISNGGYYWS3A5.278 LCDR1GGNNIGSKNVYSEQ ID NO: 4SEQ ID NO: 53A5.278 HCDR2YIYYSGSTY3A5.278 LCDR2DDSDRPSSEQ ID NO: 6SEQ ID NO: 73A5.278 HCDR3LGNWFDY3A5.278 LCDR3QVWDSSSDYVVSEQ ID NO: 8SEQ ID NO: 543A5.278 VHQVQLQESGPGLVKPSQTLSLTCTVSGGSISNGGYYWSWIRQHPGKGLEWIGYIYYSGSTYYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCASLGNWFDYWGQGTLVTVSSSEQ ID NO: 163A5.278 VLSYVLTQPPSVSVAPGQTARITCGGNNIGSKNVYWYQQKPGQAPVLVVHDH95bYDSDRPSGIPERFSGSNSGNTATLTISRVEVGDEADYSCQVWDSSSDYVVFGGGTKLTVLGSEQ ID NO: 553A5.278 HCQVQLQESGPGLVKPSQTLSLTCTVSGGSISNGGYYWSWIRQHPGKGLEWIGYIYYSGSTYYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCASLGNWFDYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGSEQ ID NO: 183A5.278 LCSYVLTQPPSVSVAPGQTARITCGGNNIGSKNVYWYQQKPGQAPVLVVHDH95bYDSDRPSGIPERFSGSNSGNTATLTISRVEVGDEADYSCQVWDSSSDYVVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECSSEQ ID NO: 56Antibody 3A5.2793A5.279 HCDR1GGSISNGGYYWS3A5.279 LCDR1GGNNIGSKNVYSEQ ID NO: 4SEQ ID NO: 53A5.279 HCDR2YIYYSGSTY3A5.279 LCDR2DDSDRPSSEQ ID NO: 6SEQ ID NO: 73A5.279 HCDR3LGNWFDY3A5.279 LCDR3QVWDSSSDDVVSEQ ID NO: 8SEQ ID NO: 573A5.279 VHQVQLQESGPGLVKPSQTLSLTCTVSGGSISNGGYYWSWIRQHPGKGLEWIGYIYYSGSTYYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCASLGNWFDYWGQGTLVTVSSSEQ ID NO: 163A5.279 VLSYVLTQPPSVSVAPGQTARITCGGNNIGSKNVYWYQQKPGQAPVLVVHDH95bDDSDRPSGIPERFSGSNSGNTATLTISRVEVGDEADYSCQVWDSSSDDVVFGGGTKLTVLGSEQ ID NO: 583A5.279 HCQVQLQESGPGLVKPSQTLSLTCTVSGGSISNGGYYWSWIRQHPGKGLEWIGYIYYSGSTYYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCASLGNWFDYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGSEQ ID NO: 183A5.279 LCSYVLTQPP SVSVAPGQTARITCGGNNIGSKNVYWYQQKPGQAPVLVVHDH95bDDSDRPSGIPERFSGSNSGNTATLTISRVEVGDEADYSCQVWDSSSDDVVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECSSEQ ID NO: 59Antibody 3A5.2943A5.294 HCDR1GGSISNGGYYWS3A5.294 LCDR1GGNNIGSKNVYSEQ ID NO: 4SEQ ID NO: 53A5.294 HCDR2YIYYSGSTY3A5.294 LCDR2DDSDRPSSEQ ID NO: 6SEQ ID NO: 73A5.294 HCDR3LGNWFDY3A5.294 LCDR3QVWDSSSDHVASEQ ID NO: 8SEQ ID NO: 603A5.294 VHQVQLQESGPGLVKPSQTLSLTCTVSGGSISNGGYYWSWIRQHPGKGLEWIGYIYYSGSTYYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCASLGNWFDYWGQGTLVTVSSSEQ ID NO: 163A5.294 VLSYVLTQPP SVSVAPGQTARITCGGNNIGSKNVYWYQQKPGQAPVLVVHDV97ADSDRPSGIPERFSGSNSGNTATLTISRVEVGDEADYSCQVWDSSSDHVAFGGGTKLTVLGSEQ ID NO: 613A5.294 HCQVQLQESGPGLVKPSQTLSLTCTVSGGSISNGGYYWSWIRQHPGKGLEWIGYIYYSGSTYYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCASLGNWFDYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGSEQ ID NO: 183A5.294 LCSYVLTQPPSVSVAPGQTARITCGGNNIGSKNVYWYQQKPGQAPVLVVHDV97ADSDRPSGIPERFSGSNSGNTATLTISRVEVGDEADYSCQVWDSSSDHVAFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECSSEQ ID NO: 62Antibody 3A5.3023A5.302 HCDR1GGSISNGGYYWS3A5.302 LCDR1GGNNIGSKNVYSEQ ID NO: 4SEQ ID NO: 53A5.302 HCDR2YIYYSGSTY3A5.302 LCDR2DDSDRPSSEQ ID NO: 6SEQ ID NO: 73A5.302 HCDR3LGNWFDY3A5.302 LCDR3QVWDSSSDHVWSEQ ID NO: 8SEQ ID NO: 633A5.302 VHQVQLQESGPGLVKPSQTLSLTCTVSGGSISNGGYYWSWIRQHPGKGLEWIGYIYYSGSTYYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCASLGNWFDYWGQGTLVTVSSSEQ ID NO: 163A5.302 VLSYVLTQPPSVSVAPGQTARITCGGNNIGSKNVYWYQQKPGQAPVLVVHDV97WDSDRPSGIPERFSGSNSGNTATLTISRVEVGDEADYSCQVWDSSSDHVWFGGGTKLTVLGSEQ ID NO: 643A5.302 HCQVQLQESGPGLVKPSQTLSLTCTVSGGSISNGGYYWSWIRQHPGKGLEWIGYIYYSGSTYYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCASLGNWFDYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGSEQ ID NO: 183A5.302 LCSYVLTQPPSVSVAPGQTARITCGGNNIGSKNVYWYQQKPGQAPVLVVHDV97WDSDRPSGIPERFSGSNSGNTATLTISRVEVGDEADYSCQVWDSSSDHVWFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECSSEQ ID NO: 653A5.0973A5.097 HCDR1GGSISNGGYYWS3A5.097 LCDR1GGNNIDSKNVYSEQ ID NO: 4SEQ ID NO: 663A5.097 HCDR2YIYYSGSTY3A5.097 LCDR2DDSDRPSSEQ ID NO: 6SEQ ID NO: 73A5.097 HCDR3LGNWFDY3A5.097 LCDR3QVWDSSSDHVVSEQ ID NO: 8SEQ ID NO: 153A5.097 VHQVQLQESGPGLVKPSQTLSLTCTVSGGSISNGGYYWSWIRQHPGKGLEWIGYIYYSGSTYYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCASLGNWFDYWGQGTLVTVSSSEQ ID NO: 163A5.097 VLSYVLTQPPSVSVAPGQTARITCGGNNIDSKNVYWYQQKPGQAPVLVVHDG29DDSDRPSGIPERFSGSNSGNTATLTISRVEVGDEADYSCQVWDSSSDHVVFGGGTKLTVLGSEQ ID NO: 673A5.097 HCQVQLQESGPGLVKPSQTLSLTCTVSGGSISNGGYYWSWIRQHPGKGLEWIGYIYYSGSTYYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCASLGNWFDYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGSEQ ID NO: 183A5.097 LCSYVLTQPPSVSVAPGQTARITCGGNNIDSKNVYWYQQKPGQAPVLVVHDG29DDSDRPSGIPERFSGSNSGNTATLTISRVEVGDEADYSCQVWDSSSDHVVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECSSEQ ID NO: 683A5.046 VHCAGGTGCAGCTGCAGGAATCTGGCCCTGGCCTGGTCAAGCCCAGCCAnucleotide GACCCTGAGCCTGACCTGTACCGTGTCCGGCGGCAGCATCAGCAACGGsequenceCGGCTACTACTGGTCCTGGATCAGACAGCACCCCGGCAAGGGCCTGG(synthetic)AATGGATCGGCTACATCTACTACAGCGGCAGCACCTACTACAACCCCAGCCTGAAGTCCAGAGTGACCATCAGCGTGGACACCAGCAAGAACCAGTTCAGCCTGAAGCTGAGCAGCGTGACAGCCGCCGACACCGCCGTGTACTACTGCGCCAGCCTGGGCAATTGGTTCGACTACTGGGGCCAGGGCACCCTCGTGACAGTGTCCTCASEQ ID NO: 693A5.046 human HCGCTAGCACCAAGGGCCCCAGCGTGTTCCCCCTGGCCCCTTGTAGCAGAconstantAGCACCAGCGAGAGCACAGCCGCCCTGGGCTGCCTGGTGAAAGACTAregionCTTCCCCGAGCCCGTCACCGTGTCCTGGAACAGCGGAGCCCTGACCAGnucleotideCGGCGTGCACACCTTTCCAGCCGTGCTGCAGAGCAGCGGCCTGTACAGsequenceCCTGAGCAGCGTGGTGACAGTGCCCTCCAGCAGCCTGGGCACCAAGA(synthetic)CCTACACCTGTAACGTGGACCACAAGCCCAGCAACACCAAGGTGGACAAGCGGGTGGAATCTAAGTACGGCCCACCCTGCCCCCCCTGCCCTGCCCCTGAATTTCTGGGCGGACCCTCCGTGTTCCTGTTCCCCCCAAAGCCCAAGGACACCCTGTATATCACTCGGGAGCCCGAAGTGACCTGCGTGGTGGTGGACGTGTCCCAGGAAGATCCCGAGGTCCAGTTCAATTGGTACGTGGACGGCGTGGAAGTGCACAACGCCAAGACCAAGCCCAGAGAGGAACAGTTCAACAGCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGACTGGCTGAACGGCAAAGAGTACAAGTGCAAAGTCTCCAACAAGGGCCTGCCCAGCTCCATCGAGAAAACCATCAGCAAGGCCAAGGGCCAGCCCCGCGAGCCTCAGGTGTACACACTGCCCCCCAGCCAGGAAGAGATGACCAAGAACCAGGTGTCCCTGACCTGTCTGGTGAAAGGCTTCTACCCCAGCGATATCGCCGTGGAATGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCTGTGCTGGACAGCGACGGCAGCTTCTTCCTGTACTCCCGGCTGACCGTGGACAAGAGCCGGTGGCAGGAAGGCAACGTCTTCAGCTGCAGCGTGATGCACGAGGCCCTGCACAACCACTACACCCAGAAGTCCCTGAGCCTGAGCCTGGGCSEQ ID NO: 703A5.046 VLAGCTACGTGCTGACCCAGCCTCCTAGCGTGTCCGTGGCCCCTGGCCAGnucleotideACCGCCAGAATCACCTGTGGCGGCAACAACATCGGCAGCAAGAACGTsequenceGTACTGGTATCAGCAGAAGCCCGGCCAGGCCCCCGTGCTGGTGGTGCA(synthetic)CGACGACAGCGACAGACCCAGCGGCATCCCCGAGCGGTTCAGCGGCAGCAACAGCGGCAATACCGCCACCCTGACCATCAGCCGGGTGGAAGTGGGCGACGAGGCCGACTACAGCTGCCAGGTCTGGGACAGCAGCAGCGACCACGTGGTGTTCGGCGGAGGCACCAAGCTGACCGTCCTAGGTSEQ ID NO: 713A5.046 human LCCAGCCCAAGGCCGCTCCCAGCGTGACCCTGTTCCCCCCAAGCAGCGAGconstantGAACTGCAGGCCAACAAGGCCACCCTGGTGTGCCTGATCAGCGACTTCregionTACCCTGGGGCCGTGACCGTGGCCTGGAAGGCCGATAGCAGCCCTGTGnucleotideAAGGCCGGCGTGGAAACCACCACCCCCTCCAAGCAGAGCAACAACAAsequenceATACGCCGCCAGCAGCTACCTGTCCCTGACCCCCGAGCAGTGGAAGTC(synthetic)CCACCGGTCCTACAGCTGCCAGGTGACACACGAGGGCAGCACCGTGGAAAAGACCGTGGCCCCCACCGAGTGCAGCSEQ ID NO: 72EMBODIMENTS

[0213] The following list of embodiments is intended to complement, rather than displace or supersede, the previous descriptions.

[0214] Embodiment 1. A human antibody molecule that immunospecifically binds to human IL-5 with an equilibrium affinity constant (KD) of at least about 40 pM as determined by surface plasmon resonance.

[0215] Embodiment 2. A human antibody molecule that immunospecifically binds to human IL-5, the antibody molecule comprising:

[0216] a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO: 6, a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO: 8, a light chain CDR1 comprising the amino acid sequence of SEQ ID NOs: 5, 21, 24, 27, 30, 33, 36, 39, or 66, a light chain CDR2 comprising the amino acid sequence of SEQ ID NOs: 7, 42, or 45, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NOs: 15, 48, 51, 54, 57, 60, or 63.

[0217] Embodiment 3. The antibody molecule of embodiment 1 or 2, wherein the antibody molecule comprises a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO: 6, a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO: 8, and

[0218] a. a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 5, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 15;

[0219] b. a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 21, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 15;

[0220] c. a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 24, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 15;

[0221] d. a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 27, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 15;

[0222] e. a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 30, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 15;

[0223] f. a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 33, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 15;

[0224] g. a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 36, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 15;

[0225] h. a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 39, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 15;

[0226] i. a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 66, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 15;

[0227] j. a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 5, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 42, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 15;

[0228] k. a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 5, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 45, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 15;

[0229] l. a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 5, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 48;

[0230] m. a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 5, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 51;

[0231] n. a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 5, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 54;

[0232] o. a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 5, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 57;

[0233] p. a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 5, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 60; or

[0234] q. a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 5, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 63;

[0235] wherein the position of the amino acid residues of the CDR is determined according to AbM.

[0236] Embodiment 4. The antibody molecule of embodiment 3, wherein the antibody molecule comprises a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO: 6, a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO: 8, a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 5, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 15.

[0237] Embodiment 5. The antibody molecule of any one of the previous embodiments, wherein the antibody molecule comprises a heavy chain variable region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 16 and

[0238] a. a light chain variable region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 17;

[0239] b. a light chain variable region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 22;

[0240] c. a light chain variable region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 25;

[0241] d. a light chain variable region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 28;

[0242] e. a light chain variable region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 31;

[0243] f. a light chain variable region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 34;

[0244] g. a light chain variable region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 37;

[0245] h. a light chain variable region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 40;

[0246] i. a light chain variable region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 43;

[0247] j. a light chain variable region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 46;

[0248] k. a light chain variable region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 49;

[0249] l. a light chain variable region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 52;

[0250] m. a light chain variable region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 55;

[0251] n. a light chain variable region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 58;

[0252] o. a light chain variable region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 61;

[0253] p. a light chain variable region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 64; or

[0254] q. a light chain variable region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 67.

[0255] Embodiment 6. The antibody molecule of any one of the previous embodiments, wherein the antibody molecule comprises a heavy chain variable region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 16 and a light chain variable region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 17.

[0256] Embodiment 7. The antibody molecule of any one of the previous embodiments, wherein the antibody molecule comprises:

[0257] a. a S228P mutation;

[0258] b. a M252Y mutation, a S254T mutation, and a T256E mutation;

[0259] c. a deletion of a heavy chain C-terminal lysine residue; or

[0260] d. any combination of a to c.

[0261] Embodiment 8. The antibody molecule of embodiment 7, wherein the antibody molecule comprises a S228P mutation and a deletion of a heavy chain C-terminal lysine residue.

[0262] Embodiment 9. The antibody molecule of embodiment 7, wherein the antibody molecule comprises a S228P mutation, a M252Y mutation, a S254T mutation, a T256E mutation, and a deletion of a heavy chain C-terminal lysine residue.

[0263] Embodiment 10. The antibody molecule of any one of the previous embodiments, wherein the antibody molecule comprises an IgG4 heavy chain constant region and a lambda light chain constant region.

[0264] Embodiment 11. The antibody molecule of any one of the previous embodiments, wherein the antibody molecule comprises a heavy chain comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 18 and

[0265] a. a light chain comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 19;

[0266] b. a light chain comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 23;

[0267] c. a light chain comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 26;

[0268] d. a light chain comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 29;

[0269] e. a light chain comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 32;

[0270] f. a light chain comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 35;

[0271] g. a light chain comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 38;

[0272] h. a light chain comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 41;

[0273] i. a light chain comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 44;

[0274] j. a light chain comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 47;

[0275] k. a light chain comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 50;

[0276] l. a light chain comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 53;

[0277] m. a light chain comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 56;

[0278] n. a light chain comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 59;

[0279] o. a light chain comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 62;

[0280] p. a light chain comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 65; or

[0281] q. a light chain comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 68.

[0282] Embodiment 12. The antibody molecule of any one of embodiments 1-10, wherein the antibody molecule comprises a heavy chain comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to

[0283] the amino acid sequence of SEQ ID NO: 20 and a light chain comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 19.

[0284] Embodiment 13. The antibody molecule of embodiment 2 or embodiment 3, wherein the antibody molecule is a Fab fragment, a Fab2 fragment, or a single chain antibody.

[0285] Embodiment 14. The antibody molecule of any one of the previous embodiments, wherein the antibody molecule has one or more of the following properties:

[0286] a. reduces binding of IL-5 to the IL-5 receptor;

[0287] b. has a serum half-life of at least about 20 days; or

[0288] c. binds human and cynomolgus monkey IL-5 but not mouse, rat, or guinea pig IL-5.

[0289] Embodiment 15. The antibody molecule of embodiment 2, wherein the antibody molecule binds to human IL-5 with an equilibrium affinity constant (KD) of at least about 40 pM as determined by surface plasmon resonance.

[0290] Embodiment 16. A pharmaceutical composition comprising the antibody molecule of any one of embodiments 1 to 15.

[0291] Embodiment 17. A nucleic acid molecule encoding the antibody molecule of any one of embodiments 1 to 15.

[0292] Embodiment 18. A vector comprising the nucleic acid molecule of embodiment 17.

[0293] Embodiment 19. A cell transformed to express the antibody molecule of any one of embodiments 1 to 15.

[0294] Embodiment 20. A method of treating a subject having eosinophilic asthma, hypereosinophilic syndrome, nasal polyposis with eosinophilic involvement, eosinophilic granulomatosis with polyangiitis, atopic dermatitis, or eosinophilic esophagitis comprising:

[0295] administering to the subject a therapeutically effective amount of the antibody molecule of any one of embodiments 1 to 15 or the pharmaceutical composition of embodiment 16 to treat the eosinophilic asthma, hypereosinophilic syndrome, nasal polyposis with eosinophilic involvement, eosinophilic granulomatosis with polyangiitis, atopic dermatitis or eosinophilic esophagitis.

[0296] Embodiment 21. Use of an effective amount of the antibody molecule of any one of embodiments 1 to 15 or the pharmaceutical composition of embodiment 16 in the treatment of eosinophilic asthma, hypereosinophilic syndrome, nasal polyposis with eosinophilic involvement, eosinophilic granulomatosis with polyangiitis, atopic dermatitis, or eosinophilic esophagitis.

[0297] Embodiment 22. Use of the antibody molecule of any one of embodiments 1 to 15 or the pharmaceutical composition of embodiment 16 in the manufacture of a medicament for the treatment of eosinophilic asthma, hypereosinophilic syndrome, nasal polyposis with eosinophilic involvement, eosinophilic granulomatosis with polyangiitis, atopic dermatitis or eosinophilic esophagitis.

Examples

examples

[0153]The following examples are provided to further describe some of the embodiments disclosed herein. The examples are intended to illustrate, not to limit, the disclosed embodiments.

Generation of Anti-Human-IL-5 Antibodies

[0154]Anti-human-IL-5 antibodies were obtained from transgenic rats (OMT) with human V-genes cloned into their genomes and which produce antibodies with human V-domains and rat Fc domains. Briefly, the transgenic rats were genetically immunized with DNA encoding IL-5 four times over 21 days (on days 0, 7, 14, 21) and boosted with recombinant human IL-5 at day 28 of the immunization protocol. Serum antibody titres were determined at days 0 and 38 of the immunization protocol by an ELISA assay using recombinant human IL-5. Briefly, sera from each animal were diluted in PBS 1% BSA and were tested using ELISA plates coated with 1 μg / ml human IL-5, or BSA as a control. A goat anti-rat IgG R-phycoerythrin conjugate (SOUTHERN BIOTECH®, #3030-09) was used at 10 μg / ml as...

embodiments

[0213]The following list of embodiments is intended to complement, rather than displace or supersede, the previous descriptions.[0214]Embodiment 1. A human antibody molecule that immunospecifically binds to human IL-5 with an equilibrium affinity constant (KD) of at least about 40 pM as determined by surface plasmon resonance.[0215]Embodiment 2. A human antibody molecule that immunospecifically binds to human IL-5, the antibody molecule comprising:[0216]a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO: 6, a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO: 8, a light chain CDR1 comprising the amino acid sequence of SEQ ID NOs: 5, 21, 24, 27, 30, 33, 36, 39, or 66, a light chain CDR2 comprising the amino acid sequence of SEQ ID NOs: 7, 42, or 45, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NOs: 15, 48, 51, 54, 57, 60, or 63.[0217]Embodiment 3. The antibody ...

CROSS REFERENCE TO RELATED APPLICATIONS

[0001] This application is a continuation of U.S. application Ser. No. 15 / 848,090, filed Dec. 20, 2017 (now U.S. Pat. No. 11,111,292), which claims priority to U.S. Provisional Application No. 62 / 438,502, filed Dec. 23, 2016, the disclosures of each of which are hereby incorporated by reference in their entirety.SEQUENCE LISTING

[0002] The instant application contains a sequence listing which has been submitted electronically in ASCII format and is hereby incorporated by reference in its entirety. Said ASCII copy, created on Jul. 26, 2021, is named 102085.001560_sl.txt and is 91 KB in size.TECHNICAL FIELD

[0003] Disclosed herein are novel antibody molecules that immunospecifically bind IL-5, and uses of the disclosed antibodies.BACKGROUND

[0004] Interleukin-5 (IL-5) is a T-helper 2 (Th2) cytokine which causes the proliferation and differentiation of both B cells and eosinophils. In B cells, IL-5 also increases immunoglobulin secretion. IL-5 is a key modulator of eosinophils, where it also regulates maturation, migration to tissues, survival, and the prevention of apoptosis.

[0005] Through two separate motifs, IL-5 binds to its specific receptor (IL5-Rα) and a signaling receptor, a common β-chain (βc) shared between Interleukin-3 (IL-3) and granulocyte-macrophage colony stimulating factor (GMCSF). The affinity of IL-5 for IL5-Rα has been reported to be in the mid-low nM range (0.2-100 nM); this shifts into the mid-pM range (˜100 pM) in the presence of βc. IL5-Rα binds to IL-5 specifically, which then recruits βc to IL-5R.

[0006] The therapeutic potential of targeting interleukin-5 (IL-5) has been demonstrated by extensive validation in the literature and recent positive Phase III clinical data for both reslizumab and mepolizumab.SUMMARY

[0007] Disclosed herein are human antibody molecules that immunospecifically bind to human IL-5 with an equilibrium affinity constant (KD) of at least about 40 pM as determined by surface plasmon resonance.

[0008] Also provided are antibody molecules comprising a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO: 6, a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO: 8, a light chain CDR1 comprising the amino acid sequence of SEQ ID NOs: 5, 21, 24, 27, 30, 33, 36, 39, or 66, a light chain CDR2 comprising the amino acid sequence of SEQ ID NOs: 7, 42, or 45, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NOs: 15, 48, 51, 54, 57, 60, or 63.

[0009] The disclosed antibody molecules can comprise a heavy chain variable region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 16 and a light chain variable region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 17, 22, 25, 28, 31, 34, 37, 40, 43, 46, 49, 52, 55, 58, 61, 64, or 67, wherein the variability (i.e. the at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identity) occurs outside of the CDR sequence.

[0010] The disclosed antibody molecules can comprise a heavy chain comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NOs: 18 or 20 and a light chain comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 19, 23, 26, 29, 32, 35, 38, 41, 44, 47, 50, 53, 56, 59, 62, 65, or 68, wherein the variability (i.e. the at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identity) occurs outside of the CDR sequence.

[0011] Nucleic acid molecules encoding the disclosed antibody molecules, vectors comprising the nucleic acid molecules, and cells transformed to express the disclosed antibody molecules are also provided.

[0012] Also disclosed are pharmaceutical compositions comprising any of the antibody molecules disclosed herein.

[0013] Also provided are methods of treating a subject having eosinophilic asthma, hypereosinophilic syndrome, nasal polyposis with eosinophilic involvement, eosinophilic granulomatosis with polyangiitis, atopic dermatitis or eosinophilic esophagitis comprising administering to the subject a therapeutically effective amount of any of the herein disclosed antibody molecules, or pharmaceutical compositions comprising the same, to treat the eosinophilic asthma, hypereosinophilic syndrome, nasal polyposis with eosinophilic involvement, eosinophilic granulomatosis with polyangiitis, atopic dermatitis or eosinophilic esophagitis.

[0014] Use of an effective amount of any of the herein disclosed antibody molecules, or pharmaceutical compositions comprising the same, in the treatment of eosinophilic asthma, hypereosinophilic syndrome, nasal polyposis with eosinophilic involvement, eosinophilic granulomatosis with polyangiitis, atopic dermatitis, or eosinophilic esophagitis are also provided.

[0015] Further provided is the use of any of the herein disclosed antibody molecules, or pharmaceutical compositions comprising the same, in the manufacture of a medicament for the treatment of eosinophilic asthma, hypereosinophilic syndrome, nasal polyposis with eosinophilic involvement, eosinophilic granulomatosis with polyangiitis, atopic dermatitis or eosinophilic esophagitis.BRIEF DESCRIPTION OF THE DRAWINGS

[0016] The summary, as well as the following detailed description, is further understood when read in conjunction with the appended drawings. For the purpose of illustrating the disclosed antibody molecules, methods, and uses, there are shown in the drawings exemplary embodiments of the antibody molecules, methods, and uses; however, the antibody molecules, methods, and uses are not limited to the specific embodiments disclosed. In the drawings:

[0017] FIG. 1 illustrates a TF-1.6G4 assay showing that antibody variant 3A5.001 (in IgG4 format) retained an equivalent potency to the original antibody 3A5 (in IgG1 format).

[0018] FIG. 2 illustrates a TF-1.6G4 assay showing that antibody variant 3A5.040 retained an equivalent potency to the parent antibody 3A5.001.

[0019] FIG. 3 is a graphical matrix showing the position and identity of the different single amino acid substitutions that were generated on the 3A5.040 VH CDRs, aligned to the original 3A5.040 VH sequence (top sequence). The boxes contain the residues which were designated CDR residues according to the AbM nomenclature. In addition to the CDR residues, variants were made to Kabat residue numbers 93 and 94 (adjacent to HC CDR3) as described in the Example “Functional testing and characterization of antibodies.” The various sequences recited in this figure are provided in SEQ ID NO:73.

[0020] FIG. 4 is graphical matrix showing the position and identity of the different single amino acid substitutions that were generated to the 3A5.040 VL CDRs, aligned to the original 3A5.040 VL sequence (top sequence). The boxes contain the residues which were designated CDR residues according to the AbM nomenclature. The various sequences recited in this figure are provided in SEQ ID NO:74.

[0021] FIG. 5 illustrates exemplary light chain CDR consensus sequences aligned to the 3A5.040 VL sequence (top sequence). 3A5.040 VL CDR single amino acid substitution variants which showed potential improvements according to the inclusion criteria described in the Examples of (the ratio of variant kd to 3A5.040 kd≥1.5) and (the ratio of variant expression level to 3A5.040 expression level ≥0.5) were included in the consensus sequences. CDR definitions and numbering are according to AbM and Kabat nomenclature, respectively. The boxes identify the positions of CDR residues. The various sequences recited in this figure are provided in SEQ ID NO:75.

[0022] FIG. 6 illustrates a multi-concentration BIACORE™ kinetic analysis of 3A5.046 antibody binding to recombinant human IL-5 at 2.5, 1.25, 0.625, 0.313 and 0.156 μg / mL. Sensorgrams show decreasing concentrations of IL-5 in order from 2.5 μg / mL IL-5 in the top trace to 0.156 μg / mL IL-5 in the bottom trace.

[0023] FIG. 7A and FIG. 7B illustrate the proliferation of TF-1.6G4 cells in response to IL-5, and the potency of inhibition of IL-5 driven proliferation by 3A5.046. 3A5.046 was a more potent inhibitor of human IL-5 (FIG. 7A) and cynomolgus monkey IL-5 (FIG. 7B) than mepolizumab.

[0024] FIG. 8A and FIG. 8B illustrate an exemplary ELISA developed using the 3A5 antibody and a control capture antibody (R&D SYSTEMS®). The ELISA was able to detect recombinant IL-5 (FIG. 8A) and IL-5 produced from CD3 / CD28 / IL-33 activated primary human T-cells from 3 donors (FIG. 8B). FIG. 8B top panel: Donor 1, FIG. 8B middle panel: Donor 3; FIG. 8B bottom panel: Donor 4.

[0025] FIG. 9 illustrates the results of an experiment in which CD34+ cord blood cells differentiated into phenotypically mature eosinophils using IL-5 and other cytokines as described in the Examples. Antibody 3A5.046 was more potent than mepolizumab at inhibiting IL-5 induced eosinophil differentiation.

[0026] FIG. 10A, FIG. 10B, FIG. 10C, FIG. 10D, and FIG. 10E illustrate the results of the BIACORE™ crossreactivity analysis of antibody 3A5.046 binding to human IL-5 (FIG. 10A), cynomolgus monkey IL-5 (FIG. 10B), mouse IL-5 (FIG. 0C), rat IL-5 (FIG. 10D), and guinea pig IL-5 (FIG. 10E). Double-referenced sensorgrams are shown for binding to cytokines at 1 μg / ml or 10 μg / ml.

[0027] FIG. 11A, FIG. 11B, FIG. 11C, and FIG. 11D illustrate the results of a BIACORE™ specificity analysis of antibody 3A5.046. Double-referenced sensorgrams are shown for binding to cytokines at 10 μg / ml (FIG. 11A and FIG. 11B) or 1 μg / ml (FIG. 11C and FIG. 11D).

[0028] FIG. 12 illustrates the results of Ascaris suum (A. suum) challenge in a cynomolgus monkey model of airway eosinophilia. At day 2 there was a substantial difference in lung (BALF) eosinophil numbers when comparing 3A5.046-treated and vehicle (placebo)-treated animals (p<0.01; Mann-Whitney test).

[0029] FIG. 13A and FIG. 13B illustrate the blood eosinophil response over 10 days following an A. suum challenge in cynomolgus monkeys which were pre-treated with 3A5.046 or a isotype-matched control antibody (“placebo”) one week before A. suum challenge (FIG. 13A). FIG. 13B illustrates further details of the blood eosinophil counts for animals treated with 3A5.046 antibody, up to 45 days post-challenge (52 days post-dose). Eosinophil counts remained below baseline for at least 45 days post-challenge following a single dose of 3A5.046 one week before challenge.

[0030] FIG. 14 illustrates BALF eosinophil levels in a human IL-5 knock-in (KI) rat model in response to a challenge with Alternaria alternata. This model may be used to test the potency of an anti-IL-5 antibody to modulate the numbers of BALF eosinophils following Alternaria challenge.DETAILED DESCRIPTION OF ILLUSTRATIVE EMBODIMENTS

[0031] The disclosed antibody molecules, methods, and uses may be understood more readily by reference to the following detailed description taken in connection with the accompanying figures, which form a part of this disclosure. It is to be understood that the disclosed antibody molecules, methods, and uses are not limited to the specific antibody molecules, methods, and uses described and / or shown herein, and that the terminology used herein is for the purpose of describing particular embodiments by way of example only and is not intended to be limiting of the claimed antibody molecules, methods, and uses.

[0032] Unless specifically stated otherwise, any description as to a possible mechanism or mode of action or reason for improvement is meant to be illustrative only, and the disclosed antibody molecules, methods, and uses are not to be constrained by the correctness or incorrectness of any such suggested mechanism or mode of action or reason for improvement.

[0033] Throughout this text, the descriptions refer to antibody molecules and methods of using said antibody molecules. Where the disclosure describes or claims a feature or embodiment associated with an antibody molecule, such a feature or embodiment is equally applicable to the methods of using said antibody molecule. Likewise, where the disclosure describes or claims a feature or embodiment associated with a method of using an antibody molecule, such a feature or embodiment is equally applicable to the antibody molecule.

[0034] Where a range of numerical values is recited or established herein, the range includes the endpoints thereof and all the individual integers and fractions within the range, and also includes each of the narrower ranges therein formed by all the various possible combinations of those endpoints and internal integers and fractions to form subgroups of the larger group of values within the stated range to the same extent as if each of those narrower ranges was explicitly recited. Where a range of numerical values is stated herein as being greater than a stated value, the range is nevertheless finite and is bounded on its upper end by a value that is operable within the context of the invention as described herein. Where a range of numerical values is stated herein as being less than a stated value, the range is nevertheless bounded on its lower end by a non-zero value. It is not intended that the scope of the invention be limited to the specific values recited when defining a range. All ranges are inclusive and combinable.

[0035] Reference to a particular numerical value includes at least that particular value, unless the context clearly dictates otherwise.

[0036] When values are expressed as approximations, by use of the antecedent “about,” it will be understood that the particular value forms another embodiment. The term “about” when used in reference to numerical ranges, cutoffs, or specific values is used to indicate that the recited values may vary by up to as much as 10% from the listed value. Thus, the term “about” is used to encompass variations of ±10% or less, variations of ±5% or less, variations of ±1% or less, variations of ±0.5% or less, or variations of ±0.1% or less from the specified value.

[0037] It is to be appreciated that certain features of the disclosed antibody molecules, methods, and uses which are, for clarity, described herein in the context of separate embodiments, may also be provided in combination in a single embodiment. Conversely, various features of the disclosed antibody molecules, methods, and uses that are, for brevity, described in the context of a single embodiment, may also be provided separately or in any subcombination.

[0038] As used herein, the singular forms “a,”“an,” and “the” include the plural.

[0039] Various terms relating to aspects of the description are used throughout the specification and claims. Such terms are to be given their ordinary meaning in the art unless otherwise indicated. Other specifically defined terms are to be construed in a manner consistent with the definitions provided herein.

[0040] The term “comprising” is intended to include, but is not necessarily limited to, examples encompassed by the terms “consisting essentially of” and “consisting of”; similarly, the term “consisting essentially of” is intended to include, but is not necessarily limited to, examples encompassed by the term “consisting of.”

[0041] The term “antibody molecule” is meant in a broad sense and includes full length immunoglobulin molecules and antigen binding fragments thereof.

[0042] Immunoglobulins can be assigned to five major classes, namely IgA, IgD, IgE, IgG, and IgM, depending on the heavy chain constant domain amino acid sequence. IgA and IgG are further sub-classified as the isotypes IgAQ1, IgA2, IgG1, IgG2, IgG3, and IgG4. Antibody light chains of any vertebrate species can be assigned to one of two clearly distinct types, namely kappa (x) and lambda (k), based on the amino acid sequences of their constant domains.

[0043] “Antigen binding fragment” refers to a portion of an immunoglobulin molecule that retains the antigen binding properties of the parental full length antibody (i.e. “antigen binding fragment thereof”). Exemplary antigen binding fragments can have: heavy chain complementarity determining regions (HCDR) 1, 2, and / or 3; light chain complementarity determining regions (LCDR) 1, 2, and / or 3; a heavy chain variable region (VH); a light chain variable region (VL); and combinations thereof. Antigen binding fragments include: a Fab fragment, a monovalent fragment consisting of the VL, VH, constant light (CL), and constant heavy 1 (CH1) domains; a F(ab)2 fragment, a bivalent fragment comprising two Fab fragments linked by a disulfide bridge at the hinge region; a Fd fragment consisting of the VH and CH1 domains; a Fv fragment consisting of the VL and VH domains of a single arm of an antibody; and a domain antibody (dAb) fragment (Ward et al., Nature 341:544-546, 1989), which consists of a VH domain or a VL domain. VH and VL domains can be engineered and linked together via a synthetic linker to form various types of single chain antibody designs where the VH / VL domains pair intramolecularly, or intermolecularly in those cases when the VH and VL domains are expressed by separate single chain antibody constructs, to form a monovalent antigen binding site, such as single chain Fv (scFv) or diabody, described for example in Int'l Pat. Pub. Nos. WO1998 / 44001, WO1988 / 01649, WO1994 / 13804, and WO1992 / 01047. These antibody fragments are obtained using techniques well known to those of skill in the art, and the fragments are screened for utility in the same manner as are full length antibodies.

[0044] The phrase “immunospecifically binds” refers to the ability of the disclosed antibody molecules to preferentially bind to its target (IL-5 in the case of anti-IL-5 antibody molecules) without preferentially binding other molecules in a sample containing a mixed population of molecules. Antibody molecules that immunospecifically bind IL-5 are substantially free of other antibodies having different antigenic specificities (e.g., an anti-IL-5 antibody is substantially free of antibodies that specifically bind antigens other than IL-5). Antibody molecules that immunospecifically bind IL-5, however, can have cross-reactivity to other antigens, such as orthologs of human IL-5, including Macacafascicularis (cynomolgus monkey) IL-5. The antibody molecules disclosed herein are able to immunospecifically bind both naturally-produced human IL-5 and to IL-5 which is recombinantly produced in mammalian or prokaryotic cells.

[0045] An antibody variable region consists of four “framework” regions interrupted by three “antigen binding sites.” The antigen binding sites are defined using various terms: (i) Complementarity Determining Regions (CDRs), three in the VH (HCDR1, HCDR2, HCDR3), and three in the VL (LCDRI, LCDR2, LCDR3) are based on sequence variability (Wu and Kabat J Exp Med 132:211-50, 1970; Kabat et al. Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md., 1991); and (ii) “Hypervariable regions” (“HVR” or “HV”), three in the VH (H1, H2, H3) and three in the VL (L1, L2, L3) refer to the regions of the antibody variable domains which are hypervariable in structure as defined by Chothia and Lesk (Chothia and Lesk Mol Biol 196:901-17, 1987). The AbM definition of CDRs is also widely used; it is a compromise between Kabat and Chothia numbering schemes and is so-called because it was used by Oxford Molecular's AbM antibody modelling software (Rees, A. R., Searle, S. M. J., Henry, A. H. and Pedersen, J. T. (1996) In Sternberg M. J. E. (ed.), Protein Structure Prediction. Oxford University Press, Oxford, 141-172). Other terms include “IMGT-CDRs” (Lefranc et al., Dev Comparat Immunol 27:55-77, 2003) and “Specificity Determining Residue Usage” (SDRU) (Almagro Mol Recognit 17:132-43, 2004). The International ImMunoGeneTics (IMGT) database (http: / / www_imgt_org) provides a standardized numbering and definition of antigen-binding sites. The correspondence between CDRs, HVs and IMGT delineations is described in Lefranc et al., Dev Comparat Immunol 27:55-77, 2003.

[0046] “Framework” or “framework sequences” are the remaining sequences of a variable region other than those defined to be antigen binding sites. Because the antigen binding sites can be defined by various terms as described above, the exact amino acid sequence of a framework depends on how the antigen-binding site was defined. “Human antibody,”“fully human antibody,” and like terms refers to an antibody having heavy and light chain variable regions in which both the framework and the antigen binding sites are derived from sequences of human origin. If the antibody contains a constant region, the constant region also is derived from sequences of human origin. A human antibody comprises heavy and / or light chain variable regions that are “derived from” sequences of human origin if the variable regions of the antibody are obtained from a system that uses human germline immunoglobulin or rearranged immunoglobulin genes. Such systems include human immunoglobulin gene libraries displayed on phage, and transgenic non-human animals such as mice carrying human immunoglobulin loci as described herein. “Human antibody” may contain amino acid differences when compared to the human germline or rearranged immunoglobulin sequences due to, for example, naturally occurring somatic mutations or intentional introduction of substitutions in the variable domain (framework and antigen binding sites), or constant domain. Typically, a “human antibody” is at least about 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical in amino acid sequence to an amino acid sequence encoded by a human germline or rearranged immunoglobulin gene. In some cases, a “human antibody” may contain consensus framework sequences derived from human framework sequence analyses, for example as described in Knappik et al., J Mol Biol 296:57-86, 2000, or synthetic HCDR3 incorporated into human immunoglobulin gene libraries displayed on phage, as described in, for example, Shi et al., J Mol Biol 397:385-96, 2010 and Int'l Pat. Pub. No. WO2009 / 085462. Antibodies in which antigen binding sites are derived from a non-human species are not included in the definition of “human antibody.”

[0047] Human antibodies, while derived from human immunoglobulin sequences, may be generated using systems such as phage display incorporating synthetic CDRs and / or synthetic frameworks, or can be subjected to in vitro mutagenesis to improve antibody properties in the variable regions or the constant regions or both, resulting in antibodies that do not naturally exist within the human antibody germline repertoire in vivo.

[0048] “Recombinant antibody” includes all antibodies that are prepared, expressed, created, or isolated by recombinant means, such as: antibodies isolated from an animal (e.g., a mouse) that is transgenic or transchromosomal for human immunoglobulin genes or a hybridoma prepared therefrom (described further below); antibodies isolated from a host cell transformed to express the antibody; antibodies isolated from a recombinant, combinatorial antibody library; and antibodies prepared, expressed, created, or isolated by any other means that involve splicing of human immunoglobulin gene sequences to other DNA sequences, or antibodies that are generated in vitro using Fab arm exchange.

[0049] “Monoclonal antibody” refers to a population of antibody molecules of a single molecular composition. A monoclonal antibody composition displays a single binding specificity and affinity for a particular epitope, or in a case of a bispecific monoclonal antibody, a dual binding specificity to two distinct epitopes. Monoclonal antibody therefore refers to an antibody population with single amino acid composition in each heavy and each light chain, except for possible well known alterations such as removal of C-terminal lysine from the antibody heavy chain. Monoclonal antibodies may have heterogeneous glycosylation within the antibody population. Monoclonal antibody may be monospecific or multispecific, or monovalent, bivalent or multivalent. A bispecific antibody is included in the term monoclonal antibody.

[0050] “Epitope” refers to a portion of an antigen to which an antibody specifically binds. Epitopes usually consist of chemically active (such as polar, non-polar, or hydrophobic) surface groupings of moieties such as amino acids or polysaccharide side chains and can have specific three-dimensional structural characteristics, as well as specific charge characteristics. An epitope can be composed of contiguous and / or discontiguous amino acids that form a conformational spatial unit. For a discontiguous epitope, amino acids from differing portions of the linear sequence of the antigen come in close proximity in 3-dimensional space through the folding of the protein molecule.

[0051] “Variant” refers to a polypeptide or a polynucleotide that differs from a reference polypeptide or a reference polynucleotide by one or more modifications for example, substitutions, insertions, or deletions. The term “mutation” as used herein is intended to mean one or more intentional substitutions which are made to a polypeptide or polynucleotide.

[0052] As used herein “90% identical to” encompasses at least 90% identical, 91% identical, 92% identical, 93% identical, 94% identical, 95% identical, 96% identical, 97% identical, 98% identical, 99% identical, or 100% identical to the reference item (e.g., a biological sequence). The current specification uses the term “% identical” to describe a number of sequences. As would be understood, the term “% identical” means that in a comparison of two sequences over the specified region the two sequences have the specified number of identical residues in the same position. The level of identity may be determined using CLUSTAL W with default parameters.

[0053] “Treat,”“treatment,” and like terms refer to both therapeutic treatment and prophylactic or preventative measures, and includes reducing the severity and / or frequency of symptoms, eliminating symptoms and / or the underlying cause of the symptoms, reducing the frequency or likelihood of symptoms and / or their underlying cause, improving or remediating damage caused, directly or indirectly, by the eosinophilic asthma, hypereosinophilic syndrome, nasal polyposis with eosinophilic involvement, eosinophilic granulomatosis with polyangiitis, atopic dermatitis or eosinophilic esophagitis. Treatment also includes prolonging survival as compared to the expected survival of a subject not receiving treatment. Subjects to be treated include those that have the condition or disorder as well as those prone to have the condition or disorder or those in which the condition or disorder is to be prevented.

[0054] As used herein, “administering to the subject” and similar terms indicate a procedure by which the disclosed antibody molecules or compositions comprising the same are injected into a patient such that target cells, tissues, or segments of the body of the subject are contacted with the disclosed antibody molecules.

[0055] The phrase “therapeutically effective amount” refers to an amount of the antibody molecules, as described herein, effective to achieve a particular biological or therapeutic result such as, but not limited to, biological or therapeutic results disclosed, described, or exemplified herein. The therapeutically effective amount may vary according to factors such as the disease state, age, sex, and weight of the individual, and the ability of the composition to cause a desired response in a subject. Exemplary indicators of a therapeutically effect amount include, for example, improved well-being of the patient, reduction of a disease symptom, arrested or slowed progression of disease symptoms, and / or absence of disease symptoms.

[0056] The following abbreviations are used herein: Alternaria alternata (Alternaria), Ascaris suum (A. suum); complementarity-determining region (CDR); heavy chain (HC); light chain (LC); heavy chain variable region (VH); light chain variable region (VL); surface plasmon resonance (SPR).Antibody Molecules

[0057] Disclosed herein are human antibody molecules that immunospecifically bind to human IL-5. The human antibody molecules can immunospecifically bind to human IL-5 with an equilibrium affinity constant (KD) of at least about 40 pM as determined by surface plasmon resonance (SPR). As used herein, “of at least about 40 pM” means that the disclosed antibodies immunospecifically bind human IL-5 with a KD of less than or equal to about 40 pM. For example, the disclosed antibodies can immunospecifically bind human IL-5 with a KD of about 40 pM, about 30 pM, about 20 pM, about 10 pM, or less than about 10 pM.

[0058] The disclosed human antibody molecules can comprise a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO: 6, a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO: 8, a light chain consensus CDR1 comprising the amino acid sequence of SEQ ID NO: 1, a light chain consensus CDR2 comprising the amino acid sequence of SEQ ID NO: 2, and a light chain consensus CDR3 comprising the amino acid sequence of SEQ ID NO: 3.

[0059] The light chain consensus CDR1 comprises the amino acid sequence of GX1X2X3X4X5X6KX7X8Y (SEQ ID NO: 1), wherein:

[0060] X1 is G or K;

[0061] X2 is N or D;

[0062] X3 is N or H;

[0063] X4 is I or A;

[0064] X5 is G or D;

[0065] X6 is S or K;

[0066] X7 is N or H; and

[0067] X8 is V or A.

[0068] In some embodiments, the light chain CDR1 amino acid sequence can comprise GGNNIGSKNVY (SEQ ID NO: 5). In some embodiments, the light chain CDR1 amino acid sequence can comprise GKNNIGSKNVY (SEQ ID NO: 21). In some embodiments, the light chain CDR1 amino acid sequence can comprise GGDNIGSKNVY (SEQ ID NO: 24). In some embodiments, the light chain CDR1 amino acid sequence can comprise GGNHIGSKNVY (SEQ ID NO: 27). In some embodiments, the light chain CDR1 amino acid sequence can comprise GGNNAGSKNVY (SEQ ID NO: 30). In some embodiments, the light chain CDR1 amino acid sequence can comprise GGNNIDSKNVY (SEQ ID NO: 66). In some embodiments, the light chain CDR1 amino acid sequence can comprise GGNNIGKKNVY (SEQ ID NO: 33). In some embodiments, the light chain CDR1 amino acid sequence can comprise GGNNIGSKHVY (SEQ ID NO: 36). In some embodiments, the light chain CDR1 amino acid sequence can comprise GGNNIGSKNAY (SEQ ID NO: 39).

[0069] The light chain CDR2 amino acid sequence comprises DDX8X9RPS (SEQ ID NO: 2), wherein:

[0070] X8 is S or L; and

[0071] X9 is D or S.

[0072] In some embodiments, the light chain CDR2 amino acid sequence can comprise DDSDRPS (SEQ ID NO: 7). In some embodiments, the light chain CDR2 amino acid sequence can comprise DDLDRPS (SEQ ID NO: 42). In some embodiments, the light chain CDR2 amino acid sequence can comprise DDSSRPS (SEQ ID NO: 45).

[0073] The light chain CDR3 amino acid sequence comprises QVWX10SSSDX11VX12 (SEQ ID NO: 3), wherein:

[0074] X10 is D or L;

[0075] X11 is H, S, Y, or D; and

[0076] X12 is V, A, or W.

[0077] In some embodiments, the light chain CDR3 amino acid sequence can comprise QVWDSSSDHVV (SEQ ID NO: 15). In some embodiments, the light chain CDR3 amino acid sequence can comprise QVWLSSSDHVV (SEQ ID NO: 48). In some embodiments, the light chain CDR3 amino acid sequence can comprise QVWDSSSDSVV (SEQ ID NO: 51). In some embodiments, the light chain CDR3 amino acid sequence can comprise QVWDSSSDYVV (SEQ ID NO: 54). In some embodiments, the light chain CDR3 amino acid sequence can comprise QVWDSSSDDVV (SEQ ID NO: 57). In some embodiments, the light chain CDR3 amino acid sequence can comprise QVWDSSSDHVA (SEQ ID NO: 60). In some embodiments, the light chain CDR3 amino acid sequence can comprise QVWDSSSDHVW (SEQ ID NO: 63).

[0078] The disclosed human antibody molecules can comprise a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO: 6, a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO: 8, a light chain CDR1 comprising the amino acid sequence of SEQ ID NOs: 5, 21, 24, 27, 30, 33, 36, 39, or 66, a light chain CDR2 comprising the amino acid sequence of SEQ ID NOs: 7, 42, or 45, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NOs: 15, 48, 51, 54, 57, 60, or 63. Exemplary antibody molecules comprise a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO: 6, a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO: 8, and

[0079] a. a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 5, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 15;

[0080] b. a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 21, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 15;

[0081] c. a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 24, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 15;

[0082] d. a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 27, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 15;

[0083] e. a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 30, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 15;

[0084] f. a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 33, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 15;

[0085] g. a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 36, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 15;

[0086] h. a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 39, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 15;

[0087] i. a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 66, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 15;

[0088] j. a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 5, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 42, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 15;

[0089] k. a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 5, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 45, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 15;

[0090] l. a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 5, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 48;

[0091] m. a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 5, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 51;

[0092] n. a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 5, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 54;

[0093] o. a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 5, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 57;

[0094] p. a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 5, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 60; or

[0095] q. a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 5, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 63;

[0096] wherein the position of the amino acid residues of the CDR is determined according to AbM.

[0097] The disclosed antibody molecules can comprise a heavy chain variable region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 16 and a light chain variable region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 17, 22, 25, 28, 31, 34, 37, 40, 43, 46, 49, 52, 55, 58, 61, 64, or 67, wherein the variability (i.e. the at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identity) occurs outside of the CDR sequence. Exemplary antibody molecules are provided in Table 1 and Table 15.

[0098] TABLE 1Antibody chain / domain composition summaryAntibodyVH protein / VL protein / HC protein / LC protein / ID(SEQ ID)(SEQ ID)(SEQ ID)(SEQ ID)3A53A5 VH3A5 VL3A5 HC3A5 LC(SEQ ID NO: 10)(SEQ ID NO: 11)(SEQ ID NO: 12)(SEQ ID NO: 13)3A5.0013A5 VH3A5 VL3A5.001 HC3A5 LC(SEQ ID NO: 10)(SEQ ID NO: 11)(SEQ ID NO: 14)(SEQ ID NO: 13)3A5.0403A5.040 VH3A5.040 VL3A5.040 HC3A5.040 LC(SEQ ID NO: 16)(SEQ ID NO: 17)(SEQ ID NO: 18)(SEQ ID NO: 19)3A5.0463A5.040 VH3A5.040 VL3A5.046 HC3A5.040 LC(SEQ ID NO: 16)(SEQ ID NO: 17)(SEQ ID NO: 20)(SEQ ID NO: 19)3A5.0633A5.040 VH3A5.040 VL +3A5.040 HC3A5.040 LC +(SEQ ID NO: 16)G25K(SEQ ID NO: 18)G25K(SEQ ID NO: 22)(SEQ ID NO: 23)3A5.0703A5.040 VH3A5.040 VL +3A5.040 HC3A5.040 LC +(SEQ ID NO: 16)N26D(SEQ ID NO: 18)N26D(SEQ ID NO: 25)(SEQ ID NO: 26)3A5.0823A5.040 VH3A5.040 VL +3A5.040 HC3A5.040 LC +(SEQ ID NO: 16)N27H(SEQ ID NO: 18)N27H(SEQ ID NO: 28)(SEQ ID NO: 29)3A5.0843A5.040 VH3A5.040 VL +3A5.040 HC3A5.040 LC +(SEQ ID NO: 16)I28A(SEQ ID NO: 18)I28A(SEQ ID NO: 31)(SEQ ID NO: 32)3A5.0973A5.040 VH3A5.040 VL +3A5.040 HC3A5.040 LC +(SEQ ID NO: 16)G29D(SEQ ID NO: 18)G29D(SEQ ID NO: 67)(SEQ ID NO: 68)3A5.1073A5.040 VH3A5.040 VL +3A5.040 HC3A5.040 LC +(SEQ ID NO: 16)S30K(SEQ ID NO: 18)S30K(SEQ ID NO: 34)(SEQ ID NO: 35)3A5.1253A5.040 VH3A5.040 VL +3A5.040 HC3A5.040 LC +(SEQ ID NO: 16)N32H(SEQ ID NO: 18)N32H(SEQ ID NO: 37)(SEQ ID NO: 38)3A5.1273A5.040 VH3A5.040 VL +3A5.040 HC3A5.040 LC +(SEQ ID NO: 16)V33A(SEQ ID NO: 18)V33A(SEQ ID NO: 40)(SEQ ID NO: 41)3A5.1613A5.040 VH3A5.040 VL +3A5.040 HC3A5.040 LC +(SEQ ID NO: 16)S52L(SEQ ID NO: 18)S52L(SEQ ID NO: 43)(SEQ ID NO: 44)3A5.1693A5.040 VH3A5.040 VL +3A5.040 HC3A5.040 LC +(SEQ ID NO: 16)D53S(SEQ ID NO: 18)D53S(SEQ ID NO: 46)(SEQ ID NO: 47)3A5.2323A5.040 VH3A5.040 VL +3A5.040 HC3A5.040 LC +(SEQ ID NO: 16)D92L(SEQ ID NO: 18)D92L(SEQ ID NO: 49)(SEQ ID NO: 50)3A5.276*3A5.040 VH3A5.040 VL +3A5.040 HC3A5.040 LC +(SEQ ID NO: 16)H95bS(SEQ ID NO: 18)H95bS(SEQ ID NO: 52)(SEQ ID NO: 53)3A5.278*3A5.040 VH3A5.040 VL +3A5.040 HC3A5.040 LC +(SEQ ID NO: 16)H95bY(SEQ ID NO: 18)H95bY(SEQ ID NO: 55)(SEQ ID NO: 56)3A5.279*3A5.040 VH3A5.040 VL +3A5.040 HC3A5.040 LC +(SEQ ID NO: 16)H95bD(SEQ ID NO: 18)H95bD(SEQ ID NO: 58)(SEQ ID NO: 59)3A5.2943A5.040 VH3A5.040 VL +3A5.040 HC3A5.040 LC +(SEQ ID NO: 16)V97A(SEQ ID NO: 18)V97A(SEQ ID NO: 61)(SEQ ID NO: 62)3A5.3023A5.040 VH3A5.040 VL +3A5.040 HC3A5.040 LC +(SEQ ID NO: 16)V97W(SEQ ID NO: 18)V97W(SEQ ID NO: 64)(SEQ ID NO: 65)*The lower case “b” in each of these sequences refers to the Kabat numbering for the CDR position. Kabat numbering allows for CDRs of variable sizes, by using alphanumeric numbering to denote amino acid insertions at certain positions. In these CDR sequences, additional amino acids were present, which were numbered as positions 95a and 95b (corresponding to Kabat positions 95A and 95B respectively). Thus, for antibody 3A5.276, for example, H95bS indicates a histidine (“H”) to serine (“S”) mutation at position 95B according to Kabat number relative to the 3A5.040 VL chain. The lower case notation is therefore used here to distinguish the Kabat number, separate from the mutation at the indicated Kabat position.

[0099] In some embodiments, the disclosed antibody molecules can comprise a heavy chain variable region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 16 and

[0100] a. a light chain variable region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 17;

[0101] b. a light chain variable region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 22;

[0102] c. a light chain variable region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 25;

[0103] d. a light chain variable region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 28;

[0104] e. a light chain variable region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 31;

[0105] f. a light chain variable region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 34;

[0106] g. a light chain variable region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 37;

[0107] h. a light chain variable region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 40;

[0108] i. a light chain variable region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 43;

[0109] j. a light chain variable region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 46;

[0110] k. a light chain variable region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 49;

[0111] l. a light chain variable region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 52;

[0112] m. a light chain variable region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 55;

[0113] n. a light chain variable region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 58;

[0114] o. a light chain variable region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 61;

[0115] p. a light chain variable region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 64; or

[0116] q. a light chain variable region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 67,

[0117] wherein the variability (i.e. the at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identity) occurs outside of the CDR sequence.

[0118] The disclosed antibody molecules can comprise one or more mutations, deletions, or insertions, in the framework and / or constant regions. In some embodiments, an IgG4 antibody molecule can comprise a S228P mutation. S228 is located in the hinge region of the IgG4 antibody molecule. Mutation of the serine (“S”) to a proline (“P”) serves to stabilize the hinge of the IgG4 and prevent Fab arm exchange in vitro and in vivo. In some embodiments, the antibody molecules can comprise one or more modifications which increase the in vivo half life of the antibody molecules. For instance in certain embodiments the antibody can comprise a M252Y mutation, a S254T mutation, and a T256E mutation (collectively referred to as the “YTE” mutation). M252, S254, and T256 are located in in the CH2 domain of the heavy chain. Mutation of these residues to tyrosine (“Y”), threonine (“T”), and glutamate (“E”), respectively, protects the antibody molecules from lysosomal degradation, thereby enhancing the serum half-life of the antibody molecules. Based on the example of other antibodies, it is contemplated that the introduction of the YTE mutation in an anti-IL-5 antibody may provide sufficient extension of serum half life to allow for administration regimes with 3 months or longer inter-dosing intervals. In some embodiments, the antibody molecules can comprise a deletion of the heavy chain C-terminal lysine residue. Deletion of the heavy chain C-terminal lysine residue reduces heterogeneity of the antibody molecules when produced by mammalian cells. In some embodiments, the antibody molecules can comprise a combination of mutations, deletions, or insertions. For example, in some aspects, the disclosed antibody molecules can comprise a S228P mutation and a deletion of a heavy chain C-terminal lysine residue. The disclosed antibodies comprising a heavy chain sequence of SEQ ID NO: 18, for example, comprise a S228P mutation and a deletion of a heavy chain C-terminal lysine residue. In some aspects, the disclosed antibody molecule can comprise a S228P mutation, a M252Y mutation, a S254T mutation, a T256E mutation, and a deletion of a heavy chain C-terminal lysine residue. The 3A5.046 antibody, for example, which comprises a heavy chain of SEQ ID NO: 20, comprises a S228P mutation, a M252Y mutation, a S254T mutation, a T256E mutation, and a deletion of a heavy chain C-terminal lysine residue.

[0119] The antibody molecule can comprise an IgG1 or IgG4 heavy chain constant region and a lambda light chain constant region. In some embodiments, the antibody molecule comprises an IgG1 heavy chain constant region and a lambda light chain constant region (antibody 3A5, for example). In some embodiments, the antibody molecule comprises an IgG4 heavy chain constant region and a lambda light chain constant region.

[0120] The disclosed antibody molecules can comprise a heavy chain comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NOs: 18 or 20 and a light chain comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 19, 23, 26, 29, 32, 35, 38, 41, 44, 47, 50, 53, 56, 59, 62, 65, or 68, wherein the variability (i.e. the at least 90% identity) occurs outside of the CDR sequence. Exemplary antibody molecules are provided in Table 1 and Table 15. In some embodiments, the antibody molecules can comprise a heavy chain comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 18 and

[0121] a. a light chain comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 19;

[0122] b. a light chain comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 23;

[0123] c. a light chain comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 26;

[0124] d. a light chain comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 29;

[0125] e. a light chain comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 32;

[0126] f. a light chain comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 35;

[0127] g. a light chain comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 38;

[0128] h. a light chain comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 41;

[0129] i. a light chain comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 44;

[0130] j. a light chain comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 47;

[0131] k. a light chain comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 50;

[0132] l. a light chain comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 53;

[0133] m. a light chain comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 56;

[0134] n. a light chain comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 59;

[0135] o. a light chain comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 62;

[0136] p. a light chain comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 65; or

[0137] q. a light chain comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 68,

[0138] wherein the variability (i.e. the at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identity) occurs outside of the CDR sequence.

[0139] The disclosed antibody molecules can comprise a heavy chain comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 20 and a light chain comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 19, wherein the variability (i.e. the at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identity) occurs outside of the CDR sequence.

[0140] In some embodiments, the antibody molecules are full length antibody molecules (with or without a deletion of the heavy chain C-terminal lysine residue). In other embodiments, the antibody molecules are antigen binding fragments. Suitable antibody binding fragments include, but are not limited to, a Fab fragment, a Fab2 fragment, or a single chain antibody.

[0141] The antibody molecules can have one or more of the following properties:

[0142] a. binds to human IL-5 with an equilibrium affinity constant (KD) of at least about 40 pM as determined by surface plasmon resonance;

[0143] b. reduces binding of IL-5 to the IL-5 receptor;

[0144] c. has a serum half-life of at least about 20 days; or

[0145] d. binds human and cynomolgus monkey IL-5 but not mouse, rat, or guinea pig IL-5.

[0146] Pharmaceutical compositions comprising any of the disclosed antibody molecules are also provided.

[0147] Also provided are nucleic acid molecules encoding any of the disclosed antibody molecules and vectors comprising the disclosed nucleic acid molecules.

[0148] Cells transformed to express any of the disclosed antibody molecules are further provided.Methods and Uses

[0149] The disclosed antibody molecules, or pharmaceutical compositions comprising the same, can be used to treat eosinophilic asthma, hypereosinophilic syndrome, nasal polyposis with eosinophilic involvement, eosinophilic granulomatosis with polyangiitis, atopic dermatitis and eosinophilic esophagitis. Any of the antibody molecule characteristics disclosed herein apply equally to the antibodies used in the disclosed methods and uses.

[0150] Disclosed herein are methods of treating a subject having eosinophilic asthma, hypereosinophilic syndrome, nasal polyposis with eosinophilic involvement, eosinophilic granulomatosis with polyangiitis, atopic dermatitis or eosinophilic esophagitis comprising administering to the subject a therapeutically effective amount of any of the antibody molecules disclosed herein, or pharmaceutical compositions comprising the same, to treat the eosinophilic asthma, hypereosinophilic syndrome, nasal polyposis with eosinophilic involvement, eosinophilic granulomatosis with polyangiitis, atopic dermatitis or eosinophilic esophagitis.

[0151] The use of an effective amount of any of the disclosed antibody molecules, or pharmaceutical compositions comprising the same, in the treatment of eosinophilic asthma, hypereosinophilic syndrome, nasal polyposis with eosinophilic involvement, eosinophilic granulomatosis with polyangiitis, atopic dermatitis or eosinophilic esophagitis is also provided.

[0152] Also provided is the use of any of the disclosed antibody molecules, or pharmaceutical compositions comprising the same, in the manufacture of a medicament for the treatment of eosinophilic asthma, hypereosinophilic syndrome, nasal polyposis with eosinophilic involvement, eosinophilic granulomatosis with polyangiitis, atopic dermatitis or eosinophilic esophagitis.EXAMPLES

[0153] The following examples are provided to further describe some of the embodiments disclosed herein. The examples are intended to illustrate, not to limit, the disclosed embodiments.Generation of Anti-Human-IL-5 Antibodies

[0154] Anti-human-IL-5 antibodies were obtained from transgenic rats (OMT) with human V-genes cloned into their genomes and which produce antibodies with human V-domains and rat Fc domains. Briefly, the transgenic rats were genetically immunized with DNA encoding IL-5 four times over 21 days (on days 0, 7, 14, 21) and boosted with recombinant human IL-5 at day 28 of the immunization protocol. Serum antibody titres were determined at days 0 and 38 of the immunization protocol by an ELISA assay using recombinant human IL-5. Briefly, sera from each animal were diluted in PBS 1% BSA and were tested using ELISA plates coated with 1 μg / ml human IL-5, or BSA as a control. A goat anti-rat IgG R-phycoerythrin conjugate (SOUTHERN BIOTECH®, #3030-09) was used at 10 μg / ml as a secondary antibody. Specific animals were chosen for hybridoma fusion based on these serum titres.

[0155] To generate hybridomas which produce monoclonal antibodies to human IL-5, splenocytes and / or lymph node cells from immunized animals were isolated and fused to P3X63Ag8.653 non-secreting mouse myeloma cells (ATCC®, CRL-1580). Cells were plated at approximately 1×105 cells / mL in flat bottom microtiter plates, followed by a two week incubation in selective medium which included 10% fetal clone serum and 1×HAT (SIGMA). Hybridomas were expanded by serial passage through four media changes in 96-well plates (96-well stages 1 to 4), then into T25 and finally T75 flasks.

[0156] The supernatants from hybridoma clones were assayed during the hybridoma expansion process, initially in a whole-cell ELISA using cells transfected with GPI-anchored human IL-5 and then ELISA using a recombinant human IL-5 ELISA assay (the latter as described above). Hybridomas which survived the scale-up process to T75 stage and which gave signals above a given threshold for binding in both of these assays were frozen as cell pellets, for cloning and sequencing of Ig v-domains. Approximately 20 hybridomas producing chimeric IgGs were selected for cloning following these screening steps.

[0157] The human v-domains from the candidate chimeric IgGs were isolated by generation of cDNA from hybridoma cell pellets, PCR amplification of v-domains, subcloning, and DNA sequencing. A total of approximately 35 heavy and light chain combinations were obtained from the sequencing of these hybridomas. All antibodies were cloned into a mammalian expression vector and transiently transfected into HEK-293 cells. Antibodies were purified using standard Protein A purification protocols.Functional Testing and Characterization of Antibodies

[0158] Selected antibodies in human IgG1 format were first assayed for specificity in a human IL-5 ELISA. Briefly, antibodies were diluted in PBS 0.1% BSA and were tested using ELISA plates coated with 1 μg / ml human IL-5, or an irrelevant control protein. A horseradish peroxidase-conjugated Anti-IgG antibody was used as a secondary antibody.

[0159] Antibodies that showed immunospecific binding to human IL-5 were ranked for potency in a human IL-5-dependent cell proliferation assay using human TF-1.6G4 (a derivative of the human erythroleukemic cell line TF-1) cells. The TF-1.6G4 cell line was subcloned and selected for enhanced surface expression of IL-5Rα and consistent proliferative response to human IL-5. The TF-1.6G4 cell line was maintained in culture following standard conditions used for the TF-1 human erythroleukemic cell line (ATCC®: CRL-2003). Briefly, dilutions of each antibody were incubated in the presence of 45 pM human IL-5 and 5×104 TF-1.6G4 cells per well, incubated for 48 h, and cell proliferation was determined using the CELLTITER-GLO® Luminescent Cell Viability Assay (PROMEGA®, WI). All proliferation and inhibition curves were fitted using a three- or four-parameter dose-response model in GraphPad Prism®6 (Version 6.04) software. Table 2 summarizes these results.

[0160] TABLE 2Summary of screening and characterizationof an initial test panel in IgG1 format.EquilibriumInhibitsSpecificityTestIC50 (pM)AffinityIL-5RαconfirmedAntibodyin TF1.6G4(KD)bindingby ELISA3A5 35.0736 pMYesYes1A3 55.518 pMYesYes2B4 56.7542 pMYesYes6G10 72.3461 pMYesYes1E8 72.4528 pMYesYes3G4 80.9241 pMYesYes6C9 81.8941 pMYesYes5H1 86.4236 pMYesYes1H10 93.2618 pMYesYes5H11109.336 pMNoYes5G9184.452 pMYesYes1B2214.445 pMYesYes

[0161] In Table 2, the results of tests were ranked in order of TF1.6G4 assay potency. The initial test panel was selected based upon potency, affinity, IL-5Rα inhibition and sequence liabilities (immunogenicity and developability).

[0162] BIACORE™ assays were used to determine the affinity of test antibodies to recombinant human IL-5 and their potency in inhibiting IL-5Rα binding to human IL-5. Table 2 summarizes these results. Binding affinity of test antibodies to human IL-5 (KD; FIG. 6) was determined on a BIACORE™ T200 system (GE HEALTHCARE®) by coating a BIACORE™ Series S Sensor Chip Protein A (GE HEALTHCARE®) with selected purified IgG antibody to a capture level of 75 RU, then recombinant human IL-5 was injected at 60 μL / min across a 7-step two-fold serial dilution range, starting at 1 μg / mL. All experiments were run using HBS-EP+ buffer (GE HEALTHCARER). The resulting sensorgrams were double-referenced (test flowcell values subtracted from control (Protein A surface with no coated antibody) flowcell values and also a buffer blank). Binding constants were determined by fitting a 1:1 Langmuir binding model to double-referenced sensorgrams.

[0163] To determine whether each test antibody inhibited the binding of IL-5 to IL-5Rα, either a BIACORE™ T200 or a BIACORE™ 3000 system (GE HEALTHCARE®) was used. A BIACORE™ CM5 Sensor Chip was first derivatized with a Fab capture kit (GE HEALTHCARER) in accordance with the manufacturer's instructions on two adjacent (test and control) flowcells. This surface was used to capture each purified IgG test antibody on a single test flowcell. Recombinant human IL-5 at 5 μg / mL or a buffer blank was then injected across both test and control flowcells, to saturate the test flowcell surface and control for non-specific association of IL-5, respectively. A second injection of purified IgG test antibody at 10 μg / mL or a buffer blank was performed on the test flowcell to block free IL-5 binding sites and control for dissociation of the IgG test antibody from the Fab capture antibody, respectively.

[0164] Subsequent injection of IL-5Rα-Fc (R&D SYSTEMS®) at either 5 or 20 μg / mL or a buffer blank across both flowcells was used to determine whether the IgG test antibody blocked the interaction between IL-5 and IL-5Rα or to control for dissociation of IgG antibody from the Fab capture antibody during this step. Antibodies which inhibited the binding of IL-5 to IL-5Rα showed a markedly reduced signal upon injection of IL-5Rα (Table 2). A triple reference subtraction method was used to analyze data in the manner detailed above. All data was exported from BIACORE™ evaluation software and subtracted in Excel (MICROSOFT®) software. All experiments were run using HBS-EP+ buffer (GE HEALTHCARE®).

[0165] A smaller panel of test antibodies (3A5, 5H11 and 2B4 in Table 2) was chosen on the basis of these results. The antibodies were reformatted as human IgG4 and tested in the same assays. The IgG4 version of antibody 3A5 (originally an IgG1) was designated as 3A5.001. This fully-human antibody was demonstrated to have equivalent potency to the original test 3A5 in IgG1 format (FIG. 1).

[0166] A variant of antibody 3A5.001 (designated 3A5.040) was prepared with specific amino acid substitutions (VH: S

[68] T, N[82A]S; VL: S[2]Y, I[3]V, Y

[92] D, wherein the residues in square brackets represent the Kabat positions) introduced in the V-domain regions to remove predicted T-cell epitopes. Antibody 3A5.040 was demonstrated to have equivalent potency to its parental antibody, 3A5.001 (FIG. 2) in the TF-1.6G4 assay.CDR Scanning of Antibody 3A5.040

[0167] Generation of antibody 3A5.040 variants—Single-mutant variants of antibody 3A5.040 were made by substituting one of a group of nine representative amino acids—A, S, L, Y, D, Q, K, H, W—at each amino acid position in the light chain CDR1 (CDR-L1), the light chain CDR2 (CDR-L2), the heavy chain CDR1 (CDR-H1) and the heavy chain CDR2 (CDR-H2) (as defined by AbM nomenclature). Antibody variants were also made by substituting one of a group of ten representative amino acids—A, S, L, Y, D, Q, K, H, W, P—at each CDR amino acid position in the light chain CDR3 (CDR-L3), heavy chain CDR3 (CDR-H3), and at Kabat positions 93 and 94 in the variable heavy chain. A complete list of all single-mutant antibody variants generated is shown in FIG. 3 (variable heavy chain) and FIG. 4 (variable light chain), respectively.

[0168] Construction of Vectors Expressing Antibodies—Variable region variants were generated by back-translation of amino acid sequences into DNA sequences which were subsequently synthesized de novo by assembly of synthetic oligonucleotides. Variable heavy (VH) variants were subcloned into a mammalian expression vector containing a human constant region to produce full-length antibody heavy chains (human IgG4 heavy chain CHI, hinge, CH2, and CH3 domains). Similarly, variable light (VL) variants were subcloned into a mammalian expression vector containing a human lambda light chain constant region to produce full-length antibody lambda chains.

[0169] Expression of antibody variants—Antibodies were produced by co-transfecting separate expression vectors encoding antibody heavy chains and light chains into EXPI293® cells (LIFE TECHNOLOGIES®, Carlsbad, CA). Each single-mutant chain was paired with a parental chain for protein expression in the EXP1293® system. For each 20 mL transfection, 3.6×107 cells were required in 20 mL of EXP1293® Expression Medium. On the day prior to transfection, cells were seeded at a density of 0.9×106 viable cells / mL and incubated overnight at 37° C. in a humidified atmosphere of 8% CO2 in air on an orbital shaker rotating at 200 rpm. On the day of transfection, the cell number and viability were determined using an automated cell counter. Only cultures with >98% viable cells were used. For each 20 mL transfection, lipid-DNA complexes were prepared by diluting 10 μg of heavy chain DNA and 10 μg of light chain DNA in OPTI-MEM® (LIFE TECHNOLOGIES®, Carlsbad, CA) I Reduced Serum Medium (Cat. no. 31985-062) to a total volume of 1.0 mL. 54 μL of EXPIFECTAMINE® 293 Reagent (LIFE TECHNOLOGIES®, Carlsbad, CA) was diluted in OPTI-MEM® I medium to a total volume of 1.0 mL. Both vials were mixed gently and incubated for 5 minutes at room temperature. Following incubation, the diluted DNA was mixed with the diluted EXPIFECTAMINE® 293 Reagent and the DNA-EXPIFECTAMINE® 293 Reagent mixture and incubated a further 20 minutes at room temperature to allow the formation of DNA-EXPIFECTAMINE® 293 Reagent complexes. Following incubation, 2 mL of DNA-EXPIFECTAMINE® 293 Reagent complex was added to each 50 mL bioreactor tube (TPP TECHNO PLASTIC PRODUCTS® AG). 2 mL of OPTI-MEM® (LIFE TECHNOLOGIES®, Carlsbad, CA) I medium was added to the negative control tube instead of DNA-EXPIFECTAMINE® 293 Reagent complex. The cells were incubated in a 37° C. incubator with a humidified atmosphere of 8% CO2 in air on an orbital shaker rotating at 200 rpm. Approximately 16-18 hours post-transfection, 100 μL of EXPIFECTAMINE® 293 Transfection Enhancer 1 and 1.0 mL of EXPIFECTAMINE® 293 Transfection Enhancer 2 were added to each bioreactor. Supernatants were harvested at approximately 48 hours post-transfection.

[0170] Purification of antibody variants—Each antibody variant was expressed in EXP1293® cells in either 20 or 100 mL of cell culture. Cultures were spun down in 50 mL falcon tubes at 3000×g for 20 minutes, and supernatants were filtered using a 0.22 μm filter. Supernatants were purified using a GILSON® ASPEC GX274 robot. Briefly, SPE cartridges (AGILENT®, 12131014) packed with 1.2 mL MABSELECT SURE® protein A resin (GE HEALTHCARE®) were pre-equilibrated with 3 column volumes of 1×PBS. Supernatant was run over the columns followed by a 4 mL 1×PBS wash. Each column was washed with 9 mL of 1 M citric acid, pH 2.9. Antibodies were eluted with 2 mL 0.1 M citric acid, pH 2.9. Antibodies were desalted into Sorensens PBS (5 mM KH2PO4, 3 mM Na2HPO4·2H2O, 145.4 mM NaCl (pH˜5.8)) using PD-10 columns (GE HEALTHCARE®).

[0171] Determination of 3A5.040 variant antibody titre in supernatant by BIACORE™—Supernatants containing antibody from expression in EXPI293® cells were analyzed using a Protein A series S chip on the BIACORE™ T200 to determine their titre and rank them with respect to the parental antibody 3A5.040. Each supernatant sample was diluted with running buffer (1×HBS-EP+, 350 mM NaCl) and captured by injection at 60 μL / min onto flow cell (FC) 2. The resulting capture level in response units (RUs) were measured on FC 2-1 by subtracting a report point 5 sec after cycle start from one 5 sec after injection. The surface was regenerated by injecting 50 mM NaOH for 12 sec at a flow rate of 60 μL / min onto FC 1 and 2 every 4 cycles. After each regeneration, the surface and sensorgram was stabilized for 120 sec by injection of running buffer. Multiple batches were run due to the large number of antibodies that were screened in this manner.

[0172] The capture levels (in BIACORE™ response units “RU”) obtained for each supernatant sample were adjusted by multiplying the values obtained for each 30 sec injection by the supernatant dilution factor, enabling comparison of capture levels between supernatants diluted to varying degrees across different experimental runs. The relative antibody expression level (titre) of each mutant was compared to a batch-specific 3A5.040 supernatant, (Table 3) using the following formula:(adjusted 3A5.040 reference RU for 30 sec injection / adjusted variant antibody RU for 30 sec injection)×100=proportional titre of parental antibody(% 3A5.040 titre)   Formula 1

[0173] Variant antibodies with a higher titre than parent 3A5.040 antibody had a “% 3A5.040 titre” value >100% and those with a lower titre, a value <100%. This enabled the identification of variant antibodies that might have improved expression over the parental antibody, 3A5.040. The titres of some variants were not determined by BIACORE™ analysis of supernatant samples, but were expressed and purified as above and their purified yields determined by spectrophotometric analysis (A280), then compared to parental 3A5.040 antibody (Table 5).

[0174] Determination of 3A5.040 variant antibody IL-5-binding kinetics by BIACORE™—Supernatants containing antibody from expression in EXPI293® cells or purified antibodies were analyzed using a Protein A series S chip on a BIACORE™ T200 system (GE HEALTHCARE®) to determine their binding affinity for recombinant human IL-5 and rank them with respect to the parental antibody 3A5.040.

[0175] Each antibody was diluted in running buffer (1×HBS-EP+, 350 mM NaCl) and captured at 60 μL / min to approximately 50 RU on FC 2. The surface and sensorgram was then stabilized for 120 sec by injection of running buffer. Recombinant human IL-5 at 5 μg / mL or running buffer was injected onto FC 1 and 2 at a flow rate of 60 μL / min for 70 sec. The IL-5 was allowed to dissociate in running buffer for 300 sec. The surface was regenerated by injecting 50 mM NaOH for 12 sec at a flow rate of 60 μL / min onto FC 1 and 2. Buffer was injected to further clean up drift for 60 sec at a flow rate of 60 μL / min onto FC 1 and 2. The surface was stabilized for 300 sec with running buffer over FC 1 and 2. Supernatant containing the parental 3A5.040 antibody and that had been transfected with each batch was run approximately every 25 cycles throughout each run. A purified sample of 3A5.040 and an IgG4 lambda isotype control was run with every batch as a measure of inter-assay variability. Multiple batches were run due to the large number of antibodies that were screened in this manner.

[0176] Data was double-referenced (flow cell 2 was subtracted from flow cell 1 and a buffer blank) and sensorgrams fitted to a 1:1 Langmuir binding model with BIACORE™ Evaluation software. A kd (off-rate) value was calculated for each 3A5.040 supernatant sample throughout the run and these were averaged to give a reference kd, with the exception of run 2.1, which only had one 3A5.040 supernatant sample (Table 3 and Tables 6-11). The calculated kd value for each supernatant antibody was compared against the 3A5.040 average reference kd using the following formula:(3A5.040 reference kd / variant antibody kd)×100=proportional kd of parental antibody(% 3A5.040 kd)   Formula 2

[0177] Variant antibodies with a lower kd (slower off-rate) than parent 3A5.040 antibody had a “% 3A5.040 kd” value >100% and those with a higher kd (faster off-rate) a value <100%. This enabled the identification of variant antibodies that might have improved binding kinetics for IL-5 and therefore improved function over the parental antibody, 3A5.040. For variants where only purified protein was made (Table 6), the BIACORE™ kinetic analysis was performed in triplicate as above using these purified antibodies and the average values from each triplicate analysis used to perform the proportional kd calculation, as above (Table 4 and Tables 8-11).

[0178] TABLE 3Determination of supematant 3A5.040 variant antibody titre and IL-5 binding kinetics by Biacore%%RunAntibodyReplicateAmino acidCaptureRmax3A5.0403A5.04#IDNumberSubstitutionlevelka (1 / Ms)kd (1 / s)KD (M)(RU)kd0 titre1.13A5.0401Wild type51.67.20E+051.15E−041.60E−1016.31.13A5.0402Wild type51.67.27E+058.96E−051.23E−1016.61.13A5.0403Wild type51.87.18E+059.21E−051.28E−1016.71.13A5.0404Wild type526.96E+059.95E−051.43E−1016.81.13A5.040AverageWild type51.757.15E+059.91E−051.39E−1016.61001001.1IgG4 54.4N / AN / AN / A0.7N / AN / AIsotype1.13A5.040Wild type56.47.17E+051.06E−041.48E−1017.893.0(Purified)1.13A5.304VH G26A54.67.41E+057.73E−051.04E−1017.1128491.13A5.305VH G26S53.17.36E+051.09E−041.48E−1016.791591.13A5.306VH G26L49.17.49E+058.74E−051.17E−1015.6113151.13A5.308VH G26D52.56.75E+058.57E−051.27E−1016.4116611.13A5.309VH G26Q55.17.47E+059.23E−051.24E−1017.4107861.13A5.310VH G26K51.87.36E+051.01E−041.37E−1016.4981001.13A5.311VH G26H50.97.60E+051.11E−041.46E−1016.389851.13A5.312VH G26W56.18.54E+051.21E−041.41E−1018821071.13A5.313VH G27A54.56.97E+051.17E−041.68E−1017.385991.13A5.314VH G27S50.97.16E+059.81E−051.37E−1016.2101951.13A5.315VH G27L54.37.01E+051.31E−041.87E−1017.1761181.13A5.316VH G27Y566.66E+058.56E−041.29E−0916.912801.13A5.317VH G27D57.46.56E+051.16E−041.76E−1017.885731.13A5.318VH G27Q55.27.31E+051.49E−042.04E−1017.467821.13A5.319VH G27K54.87.70E+052.04E−042.65E−1017.549951.13A5.320VH G27H54.47.12E+052.43E−043.42E−1016.941711.13A5.322VH S28A587.49E+051.05E−041.41E−101894621.13A5.323VH S28L53.87.18E+059.30E−051.30E−1017107701.13A5.324VH S28Y54.88.13E+058.60E−051.06E−1017115511.13A5.325VH S28D54.96.03E+059.16E−051.52E−1017108681.13A5.326VH S28Q56.96.78E+059.97E−051.47E−1017.799771.13A5.327VH S28K53.77.22E+051.15E−041.59E−1016.986841.13A5.328VH S28H547.82E+051.23E−041.57E−1016.981711.13A5.329VH S28W558.33E+051.36E−041.63E−101773411.13A5.330VH I29A56.36.54E+051.69E−042.58E−1016.859301.13A5.331VH I29S53.35.87E+052.04E−043.48E−1015.749281.13A5.332VH I29L55.38.26E+051.16E−041.41E−1017.585751.13A5.333VH I29Y55.45.44E+059.31E−041.71E−0915.711241.13A5.334VH I29D52.83.53E+065.37E−031.52E−0911.92211.13A5.336VH I29K53.76.17E+056.75E−041.10E−0914.815291.13A5.338VH I29W56.61.31E+062.34E−031.78E−0914.94381.13A5.340VH S3OL48.76.51E+051.47E−042.25E−1015.6671531.13A5.341VH S30Y54.37.54E+051.37E−041.82E−1017.3721031.13A5.342VH S3OD54.96.17E+051.23E−042.00E−1017.4811031.13A5.347VH N31A53.87.53E+056.75E−048.97E−1017.3151181.13A5.348VH N31S54.67.82E+055.03E−046.44E−1017.7201071.13A5.350VH N31Y56.29.01E+058.80E−049.76E−1017.611NE1.13A5.351VH N31D59.86.40E+052.03E−043.17E−1019.249751.13A5.352VH N31Q54.86.93E+054.21E−046.07E−1017.7241561.13A5.353VH N31K55.56.85E+057.54E−041.10E−0917.7131701.13A5.354VH N31H53.77.63E+051.32E−041.73E−1017.5751431.13A5.356VH G32A52.35.25E+053.02E−045.76E−1016.4331791.13A5.357VH G32S49.26.02E+051.23E−032.04E−0915.381531.13A5.495VH S94K556.76E+052.91E−044.31E−1017.6341271.13A5.497VH S94W56.81.67E+062.92E−031.75E−0912.53381.13A5.498VH S94P55.54.45E+051.17E−042.63E−101685171.13A5.500VH L95S2.7NENE1.13A5.504VH L95K51.71.53E+063.12E−032.03E−0911.33671.13A5.505VH L95H52.95.59E+056.66E−041.19E−0916.315881.13A5.507VH L95P54.7NB401.13A5.509VH G96S54.45.94E+051.92E−043.23E−101752881.13A5.510VH G96L544.11E+067.90E−031.92E−098.51631.13A5.513VH G96Q54.84.45E+055.13E−041.15E−0916.519641.13A5.514VH G96K54.95.14E+066.84E−031.33E−0911.411471.13A5.515VH G96H53.24.37E+051.20E−032.74E−0915.381171.13A5.516VH G96W531.31E+061.40E−021.07E−086.71551.13A5.517VH G96P47.81.35E+055.04E−043.73E−0911.720151.13A5.518VH N97A525.35E+051.72E−043.21E−1016.358951.13A5.520VH N97L54.54.73E+051.11E−042.35E−101689961.13A5.521VH N97Y54.75.05E+053.34E−046.62E−1016.130751.13A5.523VH N97Q53.75.06E+059.79E−051.94E−1016.61011271.13A5.524VH N97K52.52.89E+051.37E−044.75E−1015.1721671.13A5.525VH N97H53.14.11E+052.14E−045.20E−1015461131.13A5.526VH N97W53.75.28E+055.26E−049.95E−1015.519561.13A5.527VH N97P55.14.21E+054.38E−041.04E−0915.523681.13A5.528VH W98A57.67.26E+051.87E−042.57E−1018.4531141.13A5.529VH W98S53.96.23E+052.36E−043.78E−1016.842831.13A5.530VH W98L54.33.38E+051.72E−045.08E−1014.958381.13A5.531VH W98Y56.37.15E+051.12E−041.56E−1018.1881431.13A5.532VH W98D56.42.17E+058.87E−054.09E−1014.6112421.13A5.533VH W98Q53.85.01E+051.54E−043.08E−1015.864511.13A5.534VH W98K53.43.46E+058.25E−042.38E−0911.512731.13A5.535VH W98H53.86.13E+051.45E−042.37E−101768811.13A5.536VH W98P56.19.97E+048.17E−048.20E−0914.212311.13A5.537VH F99A54.21.54E+054.90E−043.18E−0913.720371.13A5.538VH F99S53.31.18E+051.02E−038.67E−099.910401.13A5.539VH F99L54.27.05E+051.15E−041.63E−1016.786551.13A5.540VH F99Y55.65.27E+059.13E−051.73E−1014.9109411.13A5.541VH F99D53.91.46E+051.11E−027.56E−084.41321.13A5.542VH F99Q53.89.93E+041.00E−031.01E−081310351.13A5.543VH F99K54.6NB381.13A5.544VH F99H54.82.05E+059.50E−054.64E−1014.5104411.13A5.545VH F99W54.55.35E+044.09E−047.66E−09624291.13A5.546VH F99P53.1NB431.13A5.547VH D101A55.61.68E+051.40E−048.37E−1014.771691.13A5.548VH D101S55.22.38E+051.11E−044.67E−1015.389781.13A5.549VH D101L53.65.94E+054.76E−048.01E−1016.321431.13A5.551VH D101Q54.55.93E+052.19E−043.69E−1017.145781.23A5.0401Wild type50.37.40E+051.33E−041.80E−1016.31.23A5.0402Wild type50.37.14E+058.22E−051.15E−1016.11.23A5.0403Wild type507.28E+051.01E−041.39E−1016.11.23A5.0404Wild type50.27.32E+051.10E−041.50E−1016.21.23A5.0405Wild type50.17.27E+059.36E−051.29E−1016.11.23A5.040AverageWild type50.27.28E+051.04E−041.43E−1016.21001001.2IgG450.8N / AN / AN / A0N / AIsotype1.23A5.040Wild type52.67.35E+051.11E−041.51E−1016.694(Purified)1.23A5.553VH D101H51.94.41E+051.63E−043.69E−1015.564661.23A5.555VH D101P51.37.54E+038.00E−041.06E−0723.613391.23A5.556VH Y102A50.66.60E+058.94E−051.35E−1016.41161061.23A5.557VH Y102S53.56.85E+051.11E−041.61E−1017.2941351.23A5.558VH Y102L53.27.40E+051.14E−041.54E−1016.891511.23A5.559VH Y102D52.87.03E+051.41E−042.01E−1016.774Si1.23A5.560VH Y102Q52.67.14E+051.08E−041.51E−1016.996871.23A5.561VH Y102K55.97.23E+058.01E−051.11E−1017.7130901.23A5.562VH Y102H50.47.22E+051.00E−041.39E−1016.41041321.23A5.563VH Y102W50.97.51E+051.16E−041.54E−1016.8901171.23A5.564VH Y102P536.44E+051.29E−042.01E−101581322.13A5.0401Wild type68.57.13E+051.16E−041.63E−1022.62.13A5.040AverageWild type68.57.13E+051.16E−041.63E−1022.61001002.1IgG460.7N / AN / AN / A1.6N / AIsotype2.13A5.040Wild type75.67.37E+051.05E−041.42E−1024.3110(Purified)2.13A5.048VL G24A697.16E+059.18E−051.28E−1022.71261042.13A5.049VL G24S72.56.98E+051.03E−041.47E−1023.71131042.13A5.050VL G24L73.86.79E+058.13E−051.20E−1023.91431032.13A5.051VL G24Y61.76.40E+051.12E−041.75E−1020.3104912.13A5.052VL G24D66.77.16E+059.71E−051.36E−1021.91191042.13A5.053VL G24Q72.66.99E+059.84E−051.41E−1023.6118952.13A5.054VL G24K74.27.12E+059.59E−051.35E−1024.1121982.13A5.055VL G24H60.96.90E+059.17E−051.33E−1020.2126992.13A5.056VL G24W64.46.41E+058.06E−051.26E−1021.11441112.13A5.057VL G25A69.97.04E+058.75E−051.24E−1022.91331092.13A5.059VL G25L63.17.16E+051.04E−041.45E−1021112832.13A5.060VL G25Y67.77.05E+058.55E−051.21E−1022.2136962.13A5.061VL G25D71.97.26E+051.11E−041.53E−1023.4105942.13A5.062VL G25Q73.26.52E+059.24E−051.42E−1023.5126942.13A5.064VL G25H60.46.90E+059.21E−051.33E−1020.11261082.13A5.065VL G25W70.87.37E+058.97E−051.22E−1023.1129812.13A5.066VL N26A76.87.27E+058.75E−051.20E−1024.7133772.13A5.069VL N26Y71.26.79E+058.73E−051.28E−1022.91331002.13A5.070VL N26D62.77.22E+057.87E−051.09E−1020.7147952.13A5.071VL N26Q67.47.05E+051.08E−041.53E−1022107952.13A5.072VL N26K69.77.04E+051.17E−041.66E−1022.799992.13A5.073VL N26H74.87.15E+051.11E−041.55E−1024105952.13A5.074VL N26W62.66.41E+059.82E−051.53E−1020.3118872.13A5.075VL N27A62.76.55E+059.29E−051.42E−1020.71251202.13A5.076VL N27S71.77.04E+051.09E−041.55E−1023.2106932.13A5.077VL N27L71.76.73E+058.98E−051.33E−1023.41291022.13A5.078VL N27Y61.56.16E+051.02E−041.65E−1020.31141032.13A5.080VL N27Q68.36.52E+051.07E−041.64E−1022.1108912.13A5.081VL N27K71.56.89E+058.84E−051.28E−1023.2131962.13A5.082VL N27H74.77.06E+057.85E−051.11E−1024.1148902.13A5.084VL I28A62.16.71E+056.20E−059.24E−1120.3187912.13A5.085VL I28S91.96.37E+051.16E−041.82E−1031.3100472.13A5.086VL I28L81.56.02E+051.04E−041.73E−1027.2112622.13A5.087VL I28Y73.17.07E+058.52E−051.20E−1023.8136932.13A5.088VL I28D80.35.56E+059.83E−051.77E−1027.9118402.13A5.089VL I28Q79.85.80E+051.03E−041.77E−1027.8113542.13A5.093VL G29A68.45.42E+051.10E−042.04E−1022.6105592.13A5.094VL G29S67.66.98E+059.31E−051.33E−1021.9119932.23A5.0401Wild type52.47.77E+051.05E−041.35E−1017.92.23A5.0402Wild type52.77.89E+051.09E−041.38E−1017.92.23A5.0403Wild type52.77.72E+051.37E−041.77E−1018.12.23A5.040AverageWild type52.67.79E+051.17E−041.50E−1017.91001002.2IgG456.6N / AN / AN / A0.5N / AIsotype2.23A5.040Wild type54.97.93E+051.06E−041.33E−1018.7110(Purified)2.23A5.095VL G29L55.47.17E+059.08E−051.27E−1018.7122752.23A5.096VL G29Y56.76.92E+051.33E−041.92E−1019.283582.23A5.097VL G29D56.27.31E+057.19E−059.84E−1118.7154852.23A5.099VL G29K49.97.18E+059.08E−051.26E−1017.2122712.23A5.100VL G29H527.38E+058.65E−051.17E−1017.71281002.23A5.101VL G29W55.87.59E+059.40E−051.24E−1018.71181142.23A5.102VL S30A56.36.85E+051.09E−041.58E−1018.7102842.23A5.103VL S3OL49.57.87E+059.94E−051.26E−1016.91121262.23A5.104VL S30Y54.78.17E+051.08E−041.33E−1018.5103932.23A5.105VL S3OD56.48.20E+058.91E−051.09E−1018.81251192.23A5.106VL S30Q58.37.22E+059.10E−051.26E−1019.7122782.23A5.107VL S3OK50.67.85E+057.34E−059.34E−1117.31511022.23A5.113VL K31Y53.88.35E+058.73E−051.05E−1018.4127992.23A5.114VL K31D54.54.89E+054.73E−049.65E−1017.9231052.23A5.115VL K31Q57.94.65E+054.60E−049.91E−101924852.23A5.116VL K31H49.55.52E+052.46E−044.46E−1016.7451042.23A5.117VL K31W52.75.34E+051.33E−042.49E−1017.7831252.23A5.118VL N32A55.85.07E+056.39E−041.26E−0918.317672.23A5.121VL N32Y56.19.57E+059.68E−051.01E−1019.1115912.23A5.122VL N32D49.85.68E+051.25E−042.20E−1015.8891152.23A5.123VL N32Q53.98.85E+051.25E−041.41E−1018.6891022.23A5.125VL N32H56.49.01E+057.73E−058.58E−1119.2144902.23A5.126VL N32W56.66.35E+059.26E−051.46E−1018.8120792.23A5.127VL V33A49.17.15E+058.05E−051.13E−1016.6138742.23A5.128VL V33S51.52.85E+059.65E−053.38E−1013.71151262.23A5.130VL V33Y56.17.41E+059.16E−051.24E−1019121682.23A5.131VL V33D52.47.40E+059.67E−051.31E−1019.3115402.23A5.132VL V33Q48.76.61E+051.38E−042.09E−1016.780502.23A5.135VL V33W48.45.26E+051.04E−041.99E−1018.8107242.23A5.137VL Y34S48.36.96E+059.88E−051.42E−1019112322.23A5.138VL Y34L49.45.94E+052.16E−043.64E−1016.651562.23A5.139VL Y34D52.84.63E+054.90E−041.06E−0913.123682.23A5.140VL Y34Q54.47.01E+055.40E−047.71E−1018.221552.23A5.141VL Y34K568.16E+053.72E−044.56E−1018.830642.23A5.142VL Y34H48.86.71E+055.00E−047.46E−1016.322822.23A5.143VL Y34W53.56.98E+052.19E−033.13E−099.75582.23A5.165VL S52K52.65.32E+059.30E−041.75E−0916.6121672.23A5.168VL D53A57.21.20E+062.04E−031.71E−0917.15512.23A5.172VL D53Q50.37.13E+051.08E−041.51E−1017.1103972.23A5.174VL D53H537.45E+051.23E−041.65E−1017.7901202.23A5.186VL P55S55.87.97E+051.08E−041.35E−1018.61031082.23A5.199VL S56K57.37.71E+051.18E−041.54E−1018.8941042.23A5.201VL S56W49.67.41E+051.24E−041.67E−1017.190922.23A5.210VL Q89P53.53.80E+054.78E−041.26E−091825722.23A5.211VL V90A55.77.65E+051.06E−041.39E−1018.8105783.13A5.0401Wild type53.37.72E+051.32E−041.71E−1018.23.13A5.0402Wild type53.47.76E+051.15E−041.48E−1018.13.13A5.0403Wild type53.37.73E+051.03E−041.34E−1018.13.13A5.0404Wild type547.69E+051.14E−041.49E−1018.53.13A5.0405Wild type53.87.84E+051.20E−041.53E−1018.43.13A5.040AverageWild type53.67.75E+051.17E−041.51E−1018.21001003.1IgG456N / AN / AN / A0.7N / AIsotype3.13A5.040Wild type52.58.07E+051.40E−041.74E−1017.983(Purified)3.13A5.213VL V9OL53.77.49E+051.55E−042.06E−1018.275583.13A5.214VL V90Y546.50E+052.92E−044.49E−101840853.13A5.215VL V9OD52.95.54E+054.19E−047.56E−1019.928273.13A5.216VL V90Q48.65.93E+051.88E−043.18E−1016.362743.13A5.217VL V9OK44.36.34E+058.30E−051.31E−1017141483.13A5.218VL V9OH47.55.90E+052.99E−045.07E−1017.639603.13A5.219VL V9OW53.45.69E+056.64E−041.17E−0917.918773.13A5.220VL V9OP47.72.72E+057.61E−032.79E−081.72763.13A5.221VL W91A51.83.05E+057.08E−032.32E−082.42793.13A5.223VL W91L53.62.59E+062.55E−029.83E−097.70983.13A5.224VL W91Y45.18.82E+059.03E−041.02E−0915.813643.13A5.226VL W91Q51.15.38E+063.79E−027.05E−096.80753.13A5.227VL W91K51.8NB713.13A5.228VL W91H50.91.53E+067.24E−034.74E−0992623.13A5.229VL W91P39.44.02E+057.40E−031.84E−081.42363.13A5.230VL D92A51.25.63E+059.94E−051.77E−1016.51181133.13A5.231VL D92S53.27.67E+059.46E−051.23E−1017.51241133.13A5.232VL D92L54.39.17E+057.59E−058.27E−11181541303.13A5.233VL D92Y47.71.11E+061.10E−049.92E−1116.4106723.13A5.234VL D92Q50.95.25E+051.39E−042.64E−1015.7841943.13A5.235VL 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Y34A59.91.05E+063.45E−043.28E−1017.2311704.13A5.206VL Q89D71.65.68E+058.76E−051.54E−1018.6122124.13A5.225VL W91D114.26.31E+051.05E−021.66E−083.71744.13A5.250VL S94Y74.68.14E+053.48E−044.27E−1020.3311474.13A5.252VL S94Q63.48.25E+051.65E−042.00E−1018651364.23A5.0401Wild type50.46.96E+051.03E−041.48E−1016.24.23A5.0402Wild type50.57.10E+051.06E−041.49E−10164.23A5.040AverageWild type50.47.03E+051.05E−041.49E−1016.11001004.2IgG452.5N / AN / AN / AN / AIsotype4.23A5.040Wild type48.77.21E+056.63E−059.20E−1116.5150(Purified)4.23A5.451VH G55D33.14.54E+052.50E−045.50E−109.942394.23A5.452VH G55Q37.26.09E+051.94E−043.18E−1011.754424.23A5.453VH G55K43.85.49E+052.07E−043.76E−1012.450514.23A5.454VH G55H38.25.92E+051.81E−043.05E−1011.758414.23A5.455VH G55W31.26.17E+056.65E−041.08E−091016334.23A5.456VH S56A49.76.44E+051.18E−041.84E−1015.5891044.23A5.457VH S56L51.15.49E+058.63E−051.57E−1015.61211014.23A5.458VH S56Y47.65.44E+051.92E−043.53E−1014.254854.23A5.459VH S56D495.05E+054.57E−049.06E−101523844.23A5.460VH S56Q50.25.90E+059.69E−051.64E−1015.81081064.23A5.461VH S56K52.96.11E+057.84E−051.28E−1016.61331194.23A5.463VH S56W47.74.72E+052.85E−046.03E−1013.937984.23A5.464VH T57A507.50E+30051.26E−041.67E−1016.1831044.23A5.465VH T57S48.57.29E+051.26E−041.73E−1015.683694.23A5.467VH T57Y447.66E+059.50E−051.24E−1013.8110494.23A5.468VH T57D42.57.02E+059.89E−051.41E−1014106464.23A5.469VH T57Q45.17.24E+051.07E−041.48E−1014.298584.23A5.470VH T57K46.57.28E+051.21E−041.66E−1014.686614.23A5.471VH T57H43.87.34E+051.09E−041.49E−1013.996504.23A5.472VH T57W39.27.57E+051.34E−041.77E−1013.378404.23A5.473VH Y58A48.11.56E+062.86E−031.83E−09134954.23A5.474VH Y58S50.72.76E+063.84E−031.39E−0913.731014.23A5.475VH Y58L48.65.58E+065.88E−031.05E−098.32704.23A5.476VH Y58D42.41.89E+071.68E−028.88E−107.31474.23A5.477VH Y58Q46.66.62E+051.33E−032.02E−0913.98754.23A5.478VH Y58K49.25.75E+051.31E−032.28E−0914.681024.23A5.479VH Y58H50.66.68E+055.75E−048.62E−1015.818914.23A5.480VH Y58W48.16.58E+055.53E−048.41E−1014.719794.23A5.481VH A93S45.37.18E+059.31E−051.30E−1013.9112584.23A5.482VH A93L132.55E+052.02E−047.90E−105.252174.23A5.484VH A93D0.5NENE4.23A5.485VH A93Q40.46.48E+051.00E−041.54E−1012.7105444.23A5.486VH A93K17NENE4.23A5.487VH A93H28.8NENE4.23A5.489VH A93P6.5NENE4.23A5.491VH S94L37.45.33E+058.11E−041.52E−0912.513404.23A5.492VH S94Y30.33.27E+051.10E−033.37E−099.710304.23A5.493VH S94D4.6NENE4.23A5.494VH S94Q426.74E+056.37E−049.45E−1013.616494.23A5.496VH S94H47.12.19E+064.62E−032.11E−096.52724.23A5.501VH L95Y494.17E+065.88E−031.41E−0912.42964.23A5.502VH L95D21.2NB224.23A5.506VH L95W46.46.20E+067.36E−031.19E−098.41684.23A5.511VH G96Y45.5NB594.23A5.519VH N97S485.61E+052.27E−044.04E−1014.946804.23A5.552VH D101K47.16.10E+053.63E−045.95E−1014.629674.23A5.358VH G32L48.42.01E+063.83E−031.91E−0912.33754.23A5.381VH Y34W42.47.09E+052.18E−043.07E−101448474.23A5.398VH S35bA476.82E+051.02E−041.50E−1014.7102884.23A5.488VH A93W14.2NB174.23A5.490VH S94A49.66.78E+058.70E−051.28E−1015.11201044.23A5.499VH L95A43.43.09E+067.48E−032.42E−0910.41514.23A5.508VH G96A47.15.77E+051.25E−042.17E−1014.884754.23A5.522VH N97D491.00E+061.92E−041.91E−1015.554914.23A5.554VH D101W32.71.38E+056.18E−044.47E−098.317334.23A5.144VL D50A47.15.29E+051.67E−043.15E−1014.563774.23A5.145VL DSOS48.25.74E+051.05E−041.83E−1015.1100904.23A5.146VL D5OL485.92E+052.59E−044.38E−101540864.23A5.147VL D5OY485.36E+051.95E−043.63E−101554784.23A5.148VL D50Q475.99E+051.93E−043.22E−101554704.23A5.149VL D5OK45.86.00E+051.61E−042.68E−1013.265744.23A5.150VL D5OH48.24.07E+051.68E−044.14E−1014.362954.23A5.151VL D5OW485.36E+052.35E−044.38E−101544744.23A5.152VL D51A476.34E+051.34E−042.11E−1014.678884.23A5.153VL D51S46.18.53E+051.60E−041.87E−1014.165724.23A5.154VL D51L47.86.22E+051.63E−042.63E−1014.364834.23A5.155VL D51Y48.16.14E+051.78E−042.90E−1013.759754.23A5.156VL D51Q47.86.48E+051.15E−041.77E−101591854.23A5.157VL D51K45.25.85E+051.20E−042.05E−1013.287684.23A5.158VL D51H48.36.15E+051.82E−042.95E−1014.757864.23A5.159VL D51W45.74.93E+051.96E−043.98E−1014.153574.23A5.160VL S52A47.76.61E+051.11E−041.68E−1015.294804.23A5.161VL S52L47.86.87E+055.74E−058.36E−11151821064.23A5.162VL S52Y497.26E+059.91E−051.36E−1015.81051145.13A5.0401Wild type49.96.56E+051.22E−041.86E−1014.55.13A5.0402Wild type48.56.43E+051.12E−041.75E−1014.45.13A5.0403Wild type48.16.67E+051.25E−041.87E−1013.75.13A5.0404Wild type50.46.63E+051.17E−041.77E−1014.45.13A5.040AverageWild type49.26.57E+051.19E−041.81E−1014.31001005.1IgG451.2N / AN / AN / A0.1N / AIsotype5.13A5.040Wild type48.97.07E+051.08E−041.53E−1015.6110(Purified)5.13A5.337VH I29H48.14.79E+051.12E−032.34E−0913.711245.13A5.339VH S30A50.76.79E+051.12E−041.65E−1016.310665.13A5.346VH S3OW53.46.42E+051.32E−042.06E−1016.790695.13A5.058VL G25S47.66.93E+051.08E−041.56E−1015.7110435.13A5.090VL I28K49.15.73E+051.04E−041.81E−1016.2114515.13A5.092VL I28W476.13E+051.30E−042.12E−1015.592355.13A5.212VL V9OS46.15.71E+053.11E−045.45E−1014.6384.85.13A5.251VL S94D52.92.77E+054.98E−041.79E−0915.52475.13A5.255VL S94W48.55.54E+052.86E−045.16E−1014.742195.13A5.299VL V97Q48.55.86E+059.90E−051.69E−1015.5120715.13A5.462VH S56H506.35E+051.16E−041.83E−1015.9103415.13A5.466VH T57L44.96.39E+051.02E−041.59E−1014.2117145.13A5.483VH A93Y38.2NB85.13A5.503VH L95Q48.54.04E+056.18E−041.53E−0914.419185.13A5.163VL S52D49.36.65E+051.01E−041.52E−1016.41181775.13A5.164VL S52Q49.66.57E+051.17E−041.78E−1015.81029.85.13A5.166VL S52H51.66.71E+051.11E−041.66E−1016.91071675.13A5.167VL S52W52.37.19E+051.11E−041.55E−1017.21071665.13A5.169VL D53S50.66.38E+056.35E−059.97E−1116.51871515.13A5.170VL D53L48.86.87E+059.50E−051.38E−1015.81251435.13A5.171VL D53Y516.79E+051.09E−041.60E−1016.11092085.13A5.173VL D53K52.66.35E+051.09E−041.71E−1016.71091755.13A5.175VL D53W505.88E+059.74E−051.66E−1015.71221785.13A5.176VL R54A50.46.51E+059.75E−051.50E−1016.81221385.13A5.177VL R54S50.16.45E+051.11E−041.72E−1016.81071315.13A5.178VL R54L51.56.60E+051.01E−041.53E−10171181355.13A5.179VL R54Y49.26.66E+051.24E−041.86E−1016.4961385.13A5.180VL R54D49.56.79E+059.89E−051.46E−1016.61201285.13A5.181VL R54Q51.36.58E+059.55E−051.45E−10171251485.13A5.182VL R54K51.56.50E+051.17E−041.79E−10171021655.13A5.183VL R54H49.16.47E+058.44E−051.30E−1016.31411375.13A5.184VL R54W506.14E+051.22E−041.99E−1015.8987.55.13A5.185VL P55A51.96.87E+051.34E−041.95E−1016.5898.95.13A5.187VL P55L49.67.00E+059.58E−051.37E−1016.31241535.13A5.188VL P55Y49.26.61E+051.19E−041.80E−1016.41001275.13A5.189VL P55D51.16.48E+059.21E−051.42E−1016.71291555.13A5.190VL P55Q48.86.55E+051.08E−041.64E−1016.41101515.13A5.191VL P55K49.36.38E+051.05E−041.64E−1016.31131375.13A5.192VL P55H51.56.51E+051.16E−041.78E−1016.71031765.13A5.193VL P55W51.56.41E+058.80E−051.37E−1016.81351605.13A5.194VL S56A49.86.41E+051.06E−041.65E−1016.11121435.13A5.195VL S56L49.66.85E+051.06E−041.54E−1016.31121315.13A5.196VL S56Y50.26.89E+051.14E−041.66E−1016.51041235.13A5.197VL S56D50.16.87E+051.19E−041.73E−1016.51001535.13A5.198VL S56Q49.66.86E+058.85E−051.29E−1016.41341315.13A5.200VL S56H49.36.83E+051.15E−041.68E−1016.31031385.13A5.207VL Q89K49.96.84E+051.27E−041.86E−1016.4941365.13A5.209VL Q89W48.65.57E+051.27E−042.28E−1015.794725.13A5.278VL H95bY49.97.65E+057.90E−051.03E−1016.31511395.13A5.279VL H95bD49.67.01E+057.84E−051.12E−1016.51522105.13A5.280VL H95bQ50.37.47E+051.51E−042.02E−1016.6791335.13A5.284VL V96A48.27.24E+051.88E−042.60E−1015.963655.13A5.286VL V96L48.46.50E+051.94E−042.98E−1015.961795.13A5.294VL V97A48.66.27E+057.68E−051.23E−10161551335.13A5.295VL V97S48.66.12E+051.07E−041.74E−1015.91111105.13A5.296VL V97L486.71E+059.60E−051.43E−1015.9124955.13A5.297VL V97Y48.46.72E+051.07E−041.60E−1016.2111905.13A5.298VL V97D47.75.94E+051.30E−042.18E−1015.892965.13A5.300VL V97K50.15.88E+051.18E−042.01E−1015.81014.75.13A5.301VL V97H49.46.34E+058.79E−051.39E−1016.11351685.13A5.063VL G25K496.90E+057.21E−051.05E−1016.2165905.13A5.067VL N26S496.52E+051.10E−041.69E−10161081625.13A5.083VL N27W49.66.03E+051.29E−042.14E−1016.4921485.13A5.091VL I28H47.75.71E+051.30E−042.27E−1015.792755.13A5.098VL G29Q49.76.53E+051.05E−041.61E−1016.21131395.13A5.108VL S3OH49.46.95E+051.02E−041.46E−1016.21171455.13A5.109VL S3OW486.77E+051.35E−042.00E−1015.5881125.13A5.119VL N32S48.68.35E+051.08E−041.29E−1016.3110985.13A5.120VL N32L49.28.72E+051.03E−041.18E−1016.41161195.13A5.129VL V33L48.85.80E+051.42E−042.45E−1015.884995.13A5.277VL H95bL50.46.79E+051.40E−042.06E−1016.6851805.13A5.302VL V97W47.77.18E+057.59E−051.06E−1015.9157735.13A5.303VL V97P48.76.99E+051.21E−041.73E−1016.29821This table contains a summary of the calculated kinetic and titre data for each antibody supernatant tested Variant antibodies with a lower kd (slower off-rate) than parent 3A5.040 antibody have a “% 3A5.040 kd”value >100% and those with a higher kd (faster off-rate) a value <100%. Variant antibodies with a higher titre than parent 3A5.040 antibody have a “% 3A5.040 titre”value >100% and those with a lower titre, a value <100%. N / A, non-applicable. NB, No binding. NE, No expression.

[0179] TABLE 4Determination of purified 3A5.040 variant antibody IL-5 binding kinetics by BIACORE ™%RunAmino acidCaptureRmax3A5.040#Antibody IDReplicatesSubstitutionlevelka (1 / Ms)kd (Vs)KD (M)(RU)kd63A5.040 (batch 6)1Wild type48.61.44E+062.32E−041.61E−101663A5.040 (batch 6)2Wild type49.41.44E+062.14E−041.48E−1015.763A5.040 (batch 6)3Wild type49.21.36E+062.61E−041.93E−101663A5.040 (batch 6)4Wild type49.21.36E+062.46E−041.80E−101663A5.040 (batch 6)5Wild type49.31.47E+062.52E−041.71E−1016.163A5.040 (batch 6)6Wild type49.21.45E+062.13E−041.47E−101663A5.040 (batch 6)7Wild type49.21.41E+062.29E−041.63E−1016.163A5.040 (batch 6)8Wild type50.91.40E+062.45E−041.75E−1016.863A5.040 (batch 6)AverageWild type49.31.42E+062.37E−041.67E−1016.11006IgG4 iso148.8N / AN / AN / A06IgG4 iso248.9N / AN / AN / A06IgG4 iso348.8N / AN / AN / A06IgG4 IsotypeAverage49.8N / AN / AN / A0N / A63A5.040 (Purified)1Wild type49.61.43E+062.39E−041.68E−1016.163A5.040 (Purified)2Wild type49.81.48E+062.85E−041.93E−1016.263A5.040 (Purified)3Wild type49.71.45E+062.32E−041.60E−1016.163A5.040 (Purified)AverageWild type49.71.45E+062.52E−041.74E−1016.19463A5.2371VL_D92W47.91.57E+062.67E−041.70E−1015.563A5.2372VL_D92W48.71.55E+062.58E−041.66E−1015.663A5.2373VL_D92W48.51.52E+062.12E−041.40E−1015.763A5.237AverageVL_D92W48.31.55E+062.46E−041.59E−1015.69663A5.1611VL_S52L49.41.47E+062.62E−041.78E−1016.263A5.1612VL_S52L49.91.43E+062.41E−041.69E−1016.263A5.1613VL_S52L49.91.40E+062.19E−041.57E−1016.463A5.161AverageVL_S52L49.71.43E+062.41E−041.68E−1016.29963A5.1691VL_D53S48.91.23E+062.43E−041.98E−1015.663A5.1692VL_D53S49.51.26E+062.44E−041.93E−1015.963A5.1693VL_D53S49.71.23E+062.46E−041.99E−1016.163A5.169AverageVL_D53549.31.24E+062.44E−041.97E−1015.89763A5.1831VL_R54H47.91.44E+062.32E−041.61E−1015.663A5.1832VL_R54H48.21.44E+062.50E−041.74E−1015.863A5.1833VL_R54H48.41.39E+062.04E−041.47E−1015.863A5.183AverageVL_R54H48.11.42E+062.29E−041.61E−1015.710463A5.1931VL_P55W48.51.32E+062.32E−041.76E−1015.763A5.1932VL_P55W48.31.34E+062.51E−041.88E−1015.363A5.1933VL_P55W48.71.29E+062.57E−041.99E−1015.663A5.193AverageVL_P55W48.51.32E+062.47E−041.88E−1015.59663A5.1981VL_S56Q49.11.42E+062.43E−041.71E−101663A5.1982VL_S56Q491.49E+062.90E−041.95E−1015.763A5.1983VL_S56Q49.31.42E+062.38E−041.68E−1015.963A5.198AverageVL_S56Q49.11.44E+062.57E−041.78E−1015.89263A5.2781VL_H95bY48.31.55E+062.53E−041.63E−1015.963A5.2782VL_H95bY48.21.62E+062.43E−041.49E−1015.763A5.2783VL_H95bY48.51.57E+062.14E−041.36E−1015.863A5.278AverageVL_H95bY48.31.58E+062.37E−041.49E−1015.810063A5.2791VL_H95bD501.58E+062.50E−041.58E−1016.363A5.2792VL_H95bD49.71.61E+062.42E−041.50E−1016.163A5.2793VL_H95bD501.61E+062.43E−041.51E−1016.463A5.279AverageVL_H95bD49.91.60E+062.45E−041.53E−1016.29763A5.2941VL_V97A49.11.31E+062.54E−041.94E−1015.963A5.2942VL_V97A48.61.31E+062.13E−041.62E−1015.763A5.2943VL_V97A48.61.37E+062.15E−041.57E−1015.663A5.294AverageVL_V97A48.71.33E+062.27E−041.71E−1015.710463A5.3011VL_V97H48.41.51E+062.76E−041.83E−1015.863A5.3012VL_V97H48.11.58E+061.06E−046.71E−1115.663A5.3013VL_V97H47.81.60E+062.45E−041.53E−1015.463A5.301AverageVL_V97H48.11.56E+062.09E−041.34E−1015.611463A5.0631VL_G25K481.37E+062.61E−041.90E−1015.763A5.0632VL_G25K48.11.36E+062.70E−041.99E−1015.563A5.0633VL_G25K47.81.42E+062.50E−041.76E−1015.363A5.063AverageVL_G25K47.91.38E+062.60E−041.88E−1015.59163A5.3021VL_V97W48.91.65E+062.35E−041.42E−101663A5.3022VL_V97W48.91.69E+062.35E−041.39E−1015.863A5.3023VL_V97W48.51.66E+062.40E−041.45E−1015.763A5.302AverageVL_V97W48.71.67E+062.37E−041.42E−1015.810063A5.2021VL_Q89A48.91.09E+062.49E−042.29E−1015.863A5.2022VL Q89A48.71.07E+061.89E−041.77E−1015.663A5.2023VL Q89A48.81.09E+062.44E−042.25E−1015.763A5.202AverageVL_Q89A48.81.08E+062.27E−042.10E−1015.710463A5.2031VL Q89S491.28E+062.82E−042.20E−1015.963A5.2032VL Q89S48.61.28E+062.26E−041.76E−1015.663A5.2033VL Q89S48.91.27E+062.36E−041.86E−1015.763A5.203AverageVL_Q89548.81.28E+062.48E−041.94E−1015.79663A5.2041VL Q89L49.95.01E+052.20E−044.38E−1014.463A5.2042VL Q89L49.95.02E+051.88E−043.74E−1014.263A5.2043VL Q89L49.94.95E+052.11E−044.26E−1014.363A5.204AverageVL_Q89L49.94.99E+052.06E−044.13E−1014.311563A5.2051VL Q89Y48.49.87E+052.42E−042.45E−1015.563A5.2052VL Q89Y48.61.04E+062.37E−042.28E−1015.663A5.2053VL Q89Y48.51.05E+062.60E−042.47E−1015.663A5.205AverageVL_Q89Y48.51.03E+062.46E−042.40E−1015.59663A5.2081VL_Q89H519.41E+052.53E−042.68E−101663A5.2082VL_Q89H51.19.45E+052.52E−042.66E−1016.163A5.2083VL_Q89H519.54E+052.88E−043.02E−101663A5.208AverageVL_Q89H519.47E+052.64E−042.79E−10169063A5.5501VH_D101Y48.31.16E+064.40E−043.81E−1015.563A5.5502VH_D101Y48.51.17E+064.54E−043.87E−1015.763A5.5503VH_D101Y48.31.19E+064.86E−044.10E−1015.563A5.550AverageVH_D101Y48.31.17E+064.60E−043.93E−1015.55163A5.5121VH_G96D50.41.70E+078.63E−035.07E−1013.763A5.5122VH_G96D50.71.88E+079.87E−035.26E−1013.863A5.5123VH_G96D50.51.28E+061.52E−031.19E−0912.563A5.512AverageVH_G96D50.51.24E+076.67E−037.41E−1013.33.563A5.1241VL_N32K50.99.94E+052.46E−042.47E−1016.363A5.1242VL_N32K51.21.05E+062.62E−042.49E−1016.563A5.1243VL_N32K50.71.02E+062.28E−042.24E−1016.363A5.124AverageVL_N32K50.91.02E+062.45E−042.40E−1016.39773A5.040 (batch 7)1Wild type51.21.37E+062.30E−041.68E−1016.473A5.040 (batch 7)2Wild type51.61.43E+062.15E−041.50E−1016.673A5.040 (batch 7)3Wild type51.61.42E+062.84E−042.00E−1016.673A5.040 (batch 7)AverageWild type51.41.41E+062.43E−041.73E−1016.51007IgG4 Isotype148.8N / AN / AN / A07IgG4 Isotype248.9N / AN / AN / A07IgG4 Isotype348.8N / AN / AN / A07IgG4 IsotypeAverage49.8N / AN / AN / A0N / A73A5.040 (Purified)1Wild type49.61.43E+062.39E−041.68E−1016.173A5.040 (Purified)2Wild type49.81.48E+062.85E−041.93E−1016.273A5.040 (Purified)3Wild type49.71.45E+062.32E−041.60E−1016.173A5.040 (Purified)AverageWild type49.71.45E+062.52E−041.74E−1016.19473A5.0681VL_N26L52.31.29E+062.68E−042.07E−1016.873A5.0682VL_N26L531.39E+062.08E−041.50E−1017.173A5.0683VL_N26L53.11.43E+062.69E−041.88E−1017.273A5.068AverageVL_N26L52.81.37E+062.48E−041.82E−10179873A5.0791VL_N27D49.31.22E+062.29E−041.88E−1016.173A5.0792VL_N27D501.29E+062.39E−041.85E−1016.473A5.0793VL_N27D49.91.25E+062.63E−042.09E−1016.273A5.079AverageVL_N27D49.71.25E+062.44E−041.94E−1016.2100This table contains a summary of the calculated kinetic data for each purified antibody tested. Variant antibodies with a lower kd (slower off-rate) than parent 3A5.040 antibody have a “% 3A5.040 kd” value >100% and those with a higher kd (faster off-rate) a value <100%. Runs 6 and 7 were transfected separately but run in the same BIACORE ™ batch, and therefore use the same isotype and 3A5.040 (purified) controls. N / A: non-applicable.

[0180] TABLE 5Purified protein expression yield for selected3A5.040 antibody variantsAmino acid% parent 3A5.040Run #Antibody IDSubstitutionpurified yield63A5.040Parental sequence10063A5.237VL_D92W 8963A5.161VL_S52L16063A5.169VL_D53S17363A5.183VL_R54H17263A5.193VL_P55W18763A5.198VL_S56Q20563A5.278VL_H95bY21063A5.279VL_H95bD14363A5.294VL_V97A14063A5.301VL_V97H15163A5.063VL_G25K11063A5.302VL_V97W15663A5.202VL_Q89A 6463A5.203VL_Q89S12963A5.204VL_Q89L 6163A5.205VL_Q89Y 4163A5.208VL_Q89H 5463A5.550VH_D101Y 8863A5.512VH_G96D 7263A5.124VL_N32K12873A5.040Parental Sequence10073A5.068VL_N26L 9673A5.079VL_N27D 99

[0181] TABLE 6VH CDR1 Scanning MatrixVH CDR1262728293031scGGSISNA0.5 1.31 0.80.6 0.90.3 0.60.1 1.11.2 0.1S0.6 0.91 10.3 0.51.1 0.2L0.2 1.11.2 0.80.7 1.10.8 0.91.5 0.70.7 0.2Y0.6 1.10.8 0.10.5 1.20.2 0.11 0.70.6 0.1D0.6 1.20.7 0.90.7 1.10.2 01 0.80.80.5Q0.9 1.10.8 0.70.8 10.4 0.31.2 1.11.6 0.2K1 11 0.50.8 0.90.3 0.11.6 0.81.7 0.1H0.9 0.90.7 0.40.7 0.80.2 0.10.8 0.81.4 0.8W1.1 0.81 0.20.4 0.70.4 00.7 0.90.6 0.2PVH CDR13233343535A35BscGGYYWSA1.8 0.31.2 1.31.6 0.72.900.4 0.10.9 1S1.5 0.11.201.6 0.71.300.2 0.1L0.8 00.8 0.41.6 0.61.90.11 0.10.4 0.3Y0.9 01.1 01 0.50.90D1.1 02.8 01 0.20.90NE NE0.2 0.9Q1.3 0.12.8 0.11.7 0.61.300.3 0.10.5 0.2K1.7 03.5 04.1 0.23.8NB0.3 0.10.1 0H1.1 02.7 02.2 0.1200.5 0.40.8 0W0.7 01.7 0.10.5 0.52.300.5 0PSummary of difference in 3A5.040 variant antibody titre and kd for variants of VH CDR1 (AbM definition).Kabat amino acid numbering and parent 3A5.040 amino acids are given in the top row.Mutated residue identities are listed in the left column.The leftmost value in each cell is the proportional titre of each variant vs parent 3A5.040 (>1 = higher titre, <1 = lower titre), while the right value is the proportional kd (>1 = lower kd, <1 = higher kd) of each variant vs parent 3A5.040.Variants not made are blank.NB, no binding.NE, no expression. (VH CDR1 sequence disclosed as residues 26-37 of SEQ ID NO: 73).

[0182] TABLE 7VH CDR2 Scanning MatrixVH CDR2505152535455565758scYIYYSGSTYA2.70.22.40.8302.50.21.10.60.70.910.910.810S3.30.21.30.83.30.12.30.60.70.70.70.810L1.20.30.90.72.1010.21.600.70.111.20.11.20.70Y1.100.900.50.10.90.50.51.1D1.70.10.50.13.3NB1.80.2200.40.40.80.20.51.10.50Q2.40.210.22.602.10.21.50.10.40.51.11.10.610.80.1K1.40.21.20.13.5NB2.30.1200.50.51.21.30.60.910.1H30.10.902.90.32.10.510.20.40.60.410.510.90.2W1.30.11.4010.11.50.41.10.10.30.210.40.40.80.80.2PSummary of difference in 3A5.040 variant antibody titre and kd for variants of VH CDR2 (AbM definition). Kabat amino acid numbering and parent 3A5.040 amino acids are given in the top row. Mutated residue identities are listed in the left column. The leftmost value in each cell is the proportional titre of each variant vs parent 3A5.040 (>1 = higher titre, <1 = lower titre), while the right value is the proportional kd (>1 = lower kd, <1 = higher kd) of each variant vs parent 3A5.040. Variants not made are blank. NB, no binding. NE, no expression. (VH CDR2 sequence disclosed as residues 52-60 of SEQ ID NO: 73).

[0183] TABLE 8VH CDR3 Scanning MatrixVH CDR393949596979899101102scASLGNWFDYA11.20.500.80.810.61.10.50.40.20.70.71.11.2S0.61.1NE NE0.90.50.80.50.80.40.40.10.80.91.40.9L0.20.50.40.10.6010.90.40.60.60.90.40.20.50.9Y0.1NB0.30.1100.6NB0.80.31.40.90.41.10.90.5DNENENENE0.2NB0.700.90.50.41.10.300.50.7Q0.410.50.20.20.20.60.21.310.50.60.40.10.80.50.91K0.2NB1.30.30.701.501.70.70.70.10.4 NB0.70.30.91.3H0.3NB0.700.90.11.20.11.10.50.80.70.410.70.61.31W0.2NB0.40.00.700.600.60.20.30.20.30.21.20.9P0.1NB0.20.80.4NB0.20.20.70.20.30.10.4 NB0.40.10.30.8Summary of difference in 3A5.040 variant antibody titre and kd for variants of VH CDR3 (AbM definition). and Kabat amino acid positions 93 and 94. Kabat amino acid numbering and parent 3A5.040 amino acids are given in the top row. Mutated residue identities are listed in the left column. The leftmost value in each cell is the proportional titre of each variant vs parent 3A5.040 (>1 = higher titre, <1 = lower titre), while the right value is the proportional kd (>1 = lower kd, <1 = higher kd) of each variant vs parent 3A5.040. Variants not made are blank. NB, no binding. NE, no expression. (VH CDR3 sequence disclosed as residues 98-106 of SEQ ID NO: 73).

[0184] TABLE 9VL CDR1 Scanning MatrixVL CDR1242526272829scGGNNIGA1 1.31.1 1.30.8 1.31.21.20.9 1.90.6 1.1S11.10.4 1.11.6 1.10.91.10.5 10.9 1.2L1 1.40.81.11 111.30.6 1.10.8 1.3Y0.9 11 1.41 1.311.10.9 1.40.6 0.9D1 1.20.9 11 1.5110.4 1.20.9 1.6Q1 1.20.9 1.31 1.10.91.10.5 1.11.4 1.1K1 1.20.9 1.71 111.30.5 1.10.7 1.3H1 1.31.1 1.31 10.91.50.8 0.91 1.4W1.1 1.40.8 1.30.9 1.21.50.90.4 0.91.1 1.2PVL CDR13031323334scSKNVYA0.8 1.11.20.60.7 0.20.7 1.51.70.3S1.1 0.81 1.11.3 1.20.3 1.2L1.3 1.21.3 0.31.2 1.21 0.80.6 0.5Y0.9 1.11 1.30.91.20.7 1.3D1.2 1.31.1 0.21.2 0.90.4 1.20.7 0.2Q0.8 1.30.9 0.310.90.5 0.80.6 0.2K1 1.61.3 10.1 10.6 0.3H1.5 1.21 0.50.91.50.5 NB0.8 0.2W1.1 0.91.3 0.90.81.30.2 1.10.6 0.1PSummary of difference in 3A5.040 variant antibody titre and kd for variants of VL CDRI (AbM definition).Kabat amino acid numbering and parent 3A5.040 amino acids are given in the top row.Mutated residue identities are listed in the left column. The leftmost value in each cell is the proportional titre of each variant vs parent 3A5.040 (>1 = higher titre, <1 = lower titre), while the right value is the proportional kd (>1 = lower kd, <1 = higher kd) of each variant vs parent 3A5.040.Variants not made are blank.NB, no binding.NE, no expression. (VL CDR1 sequence disclosed as residues 23-33 of SEQ ID NO: 74).

[0185] TABLE 10VL CDR2 Scanning MatrixVL CDR250515253545556scDDSDRPSA0.80.60.90.80.80.90.50.11.41.20.10.91.41.1S0.910.70.71.51.91.31.11.11L0.90.40.80.61.11.81.41.31.41.21.51.21.31.1Y0.80.50.80.61.11.12.11.11.41.01.311.21D1.81.21.31.21.61.31.51Q0.70.50.90.90.11111.51.21.51.11.31.3K0.70.60.70.91.70.11.81.11.71.01.41.110.9H10.60.90.61.71.11.20.91.41.41.811.41W0.70.40.60.51.71.11.81.20.11.01.61.40.90.9PSummary of difference in 3A5.040 variant antibody titre and kd for variants of VL CDR2 (AbM definition). Kabat amino acid numbering and parent 3A5.040 amino acids are given in the top row. Mutated residue identities are listed in the left column. The leftmost value in each cell is the proportional titre of each variant vs parent 3A5.040 (>1 = higher titre, <1 = lower titre), while the right value is the proportional kd (>1 = lower kd, <1 = higher kd) of each variant vs parent 3A5.040. Variants not made are blank. NB, no binding. NE, no expression. (VL CDR2 sequence disclosed as residues 49-55 of SEQ ID NO: 74).

[0186] TABLE 11VL CDR3 Scanning MatrixVL CDR3899091929394scQVWDSSA0.610.810.801.11.21.1011S1.3100.4101.11.2L0.61.20.60.8101.31.51.200.90.2Y0.410.90.40.60.10.71.11.201.50.3D0.11.20.30.30.700.700.10.2Q0.80.60.801.90.81.101.40.6K1.40.90.51.40.7NB0.91.31.6NB1.10.8H0.50.90.60.40.601.21.11.3010.8W0.70.90.80.2NENE1.200.20.4P0.70.20.800.400.60.81.101.40.9VL CDR39595A95B9697scSDHVV111.100.20.70.7 0.61.31.510.10.61.60.2 0.51.1 1.1110.701.80.90.8 0.61 1.20.81.1101.41.50.6 0.10.91.11.10.92.11.50.1 0.11 0.90.61.31.101.30.80.3 0.20.7 1.21.21.20.700.61.20.2 00111.21.900.5 NB1.7 1.40.90.90.901.510.200.71.61.501.501.501.1 0.20.21Summary of difference in 3A5.040 variant antibody titre and kd for variants of VL CDR3 (AbM definition).Kabat amino acid numbering and parent 3A5.040 amino acids are given in the top row.Mutated residue identities are listed in the left column.The leftmost value in each cell is the proportional titre of each variant vs parent 3A5.040 (>1 = higher titre, <1 = lower titre), while the right value is the proportional kd (>1 = lower kd, <1 = higher kd) of each variant vs parent 3A5.040.Variants not made are blank.NB, no binding.NE, no expression. (VL CDR3 sequence disclosed as residues 88-98 of SEQ ID NO: 74).Expression Vector Construction

[0187] Nucleotide sequence can influence gene expression and subsequent protein expression level. Several different nucleotide sequences were examined for optimal expression of 3A5.046. These sequences are summarized in the following table:

[0188] TABLE 12Nucleotide sequenceGene NameSEQ ID Nos:3A5.046 VHSEQ ID NO: 693A5.046 VLSEQ ID NO: 713A5.046 HC constant regionSEQ ID NO: 703A5.046 LC constant regionSEQ ID NO: 72Measurement of the Affinity of Antibody 3A5.046 for Human Recombinant IL-5 by SPR

[0189] The affinity of antibody 3A5.046 for recombinant human IL-5 was determined using a BIACORE™ T200 system (GE HEALTHCARE®). Briefly, a commercial mouse anti-human IgG antibody (THERMO SCIENTIFIC®) was coupled to two adjacent (control and test) flowcells on a BIACORE™ CM5 chip (GE HEALTHCARE®) according to the manufacturer's recommendations. The 3A5.046 antibody was captured at a low density (approximately 125 RU) on the test flowcell surface and dilutions of recombinant human IL-5 or a buffer blank were injected over both test and control flowcells, at 30 μL / min (FIG. 6, Table 13).

[0190] The resulting sensorgrams were double-referenced (test flowcell values subtracted from control (anti-human IgG-coupled surface with no coated antibody) flowcell values and also a buffer blank). Subtracted sensorgrams from injections of human IL-5 at 2.5, 1.25, 0.625, 0.313, and 0.156 μg / mL were fitted to a 1:1 Langmuir binding model using BIACORE™ Evaluation software to determine binding constants from duplicate assays (Table 13).

[0191] TABLE 13Kinetic constants from a multi-concentration BIACORE ™ kineticanalysis of 3A5.046 antibody binding to recombinanthuman IL-5 at 2.5, 1.25, 0.625, 0.313 and 0.156 μg / mLAntibodyExperimentka (1 / Ms)kd (1 / s)KD (pM)Chi2 (RU2)3A5.046Repeat 18.16E+051.58E−0519.40.0269Repeat 28.49E+051.84E−0521.70.0296

[0192] These results demonstrate that antibody 3A5.046 has a high affinity for recombinant human IL-5.Potency of 3A5.046 Compared to Mepolizumab in the TF-1.6G4 Cell Line Proliferation Assay

[0193] 3A5.046 and mepolizumab were run in the TF-1.6G4 cell line proliferation assay. 3A5.046 and mepolizumab inhibited human IL-5 proliferation with a mean IC50 of 13.8 pM and 534 pM respectively (See Table 14 and FIG. 7A). Both antibodies demonstrated inhibition of cynomolgus monkey IL-5 on the TF1.6G4 cell line (FIG. 7B), with a mean IC50 for 3A5.046 of 43.8 pM and for mepolizumab of 584 pM.

[0194] TABLE 14Number ofMeanStd.AntibodyAgonistObservations(pM)Dev.MepolizumabCynomolgus4584.0982129.9019IL-5Human IL-54534.2803150.25463A5.046Cynomolgus8 43.80916 12.62095IL-5Human IL-58 13.83667  4.219783Detection of Native IL5 from Primary Cells Via ELISA Using Anti-IL-5 Antibodies of the Invention

[0195] Antibody 3A5 was used as a capture reagent in a sandwich ELISA. Paired with a commercially available detection antibody (TRFk5R; R&D SYSTEMS®, #MAB405) quantification of recombinant IL-5 levels in spiked buffer samples could be determined (FIG. 8). This method was then applied to the detection of native IL-5 secreted by primary cells. CD4+ T-cells from four donors were stimulated to secrete IL-5. The IL-5 levels were then determined using the sandwich ELISA with antibody 3A5 as the capture reagent. Three out of four donor T-cells responded to T-cell stimulation and had detectable IL-5 levels in the cell culture supernatant (FIG. 8). Similar studies were performed on human IL-5 secreted from primary human donor CD4+ T-cells using antibody 3A5.046 as a capture agent, and the results obtained were consistent with the results obtained with antibody 3A5 (data not shown).Anti-IL-5 Antibody Inhibition of Eosinophil Differentiation

[0196] A survival assay using CD34+ cord blood cells was used to assess the capacity of anti-IL-5 antibodies to inhibit the IL-5 dependent differentiation of bone marrow progenitors to eosinophils and to assess the capacity of anti-IL-5 antibodies to inhibit phenotypically mature eosinophil survival. In IL-5 treated control cells, phenotypically mature eosinophils were the predominant population at day 28 (FIG. 9, shaded region in graph). Antibody 3A5.046 was able to inhibit the differentiation of CD34+ cord blood cells, incubated with IL-5 for 28-days, into eosinophils. Antibody 3A5.046 was a more potent inhibitor of IL-5 mediated eosinophil differentiation than mepolizumab (FIG. 9).X-Ray Crystallography Analysis

[0197] The molecular structure of the complex between antibody 3A5.046 Fab and recombinant human IL-5 is investigated by X-ray crystallography. To prepare for X-ray crystallography experiments, 3A5.046 is digested by papain, which cleaves the intact antibody in the hinge region, separating the Fc and Fab domains. The 3A5.046 Fab is purified by standard Protein A affinity chromatography and complexed with recombinant non-glycosylated human IL-5. The 3A5.046 Fab:human IL-5 complex is purified for crystallization experiments using standard size-exclusion chromatography. The complex is set up in variable concentrations, into commercially available sparse-matrix crystallization screens for broad sampling of crystallization conditions. Optimisations of any ‘hit’ conditions that provide crystals of the complex from the sparse-matrix screens are performed to improve upon crystal morphology and crystal diffraction as required.

[0198] Crystals of the 3A5.046 Fab:human IL-5 complex are harvested, flash-frozen in liquid nitrogen and transported to a synchrotron facility for diffraction tests and data collection. Crystal diffraction testing is performed at a synchrotron, which houses high energy X-ray micro beam lines for producing high-resolution diffraction. In this way, X-ray diffraction data on crystals of 3A5.046 Fab:human IL-5 complex is collected. An initial structure model of the complex may be obtained using standard molecular replacement techniques with published molecular structures of IL-5 and Fab as templates. The final structure of the 3A5.046 Fab and human IL-5 complex may be obtained by iterative refinement of the structure model, until the errors of fitting are better than 30% (final Rfree of 0.3) and chemical bonding parameters (e.g. the distribution of bond lengths and angles, stereochemistry and hydrogen bonds) are within acceptable limits.Characterisation of Antibody 3A5.046 Binding to IL-5 from Non-Human Model Species

[0199] Antibody 3A5.046 was tested by BIACORE™ assay using a T200 system (GE HEALTHCARE®) for binding to recombinant cytokines human IL-5, rat IL-5, mouse IL-5, and guinea pig IL-5. This panel of non-human IL-5s was chosen to represent major animal model species used in preclinical development. An isotype control antibody (IgG4) and antibody 3A5.046 were immobilized on flowcells 1 and 2 of a Series S CM5 chip (GE HEALTHCARE®), respectively, using standard amine coupling methods. Cytokines were diluted to 1 and 10 μg / ml in HBS-EP+ running buffer (GE HEALTHCARE®) and injected across flowcells 1 and 2. Buffer-only injections across flowcells 1 and 2 were included for double-referencing. Data analysis was performed using BIACORE™ T200 evaluation software. The resulting sensorgrams were double-referenced (test flowcell values subtracted from control flowcell values and also a buffer blank).

[0200] Antibody 3A5.046 was found to bind to human IL-5 and cynomolgus monkey IL-5, but not to mouse IL-5, rat IL-5, or guinea pig IL-5 (FIG. 10A-FIG. 10E). This indicated that 3A5.046 could be used in cynomolgus macaque preclinical models.Antibody 3A5.046 does not Bind to Other Human Cytokines Closely Related to IL-5

[0201] Antibody 3A5.046 was tested by BIACORE™ assay using a T200 system (GE HEALTHCARE®) for binding to recombinant human cytokines IL-2, IL-3, IL-4, IL-5, granulocyte macrophage colony stimulating factor (GM-CSF), macrophage colony stimulating factor (M-CSF), and growth hormone (GH). This representative panel of closely-related human cytokines was chosen based on their structural and functional similarity to human IL-5. An isotype control antibody (IgG4) and antibody 3A5.046 were immobilized on flowcells 1 and 2 of a Series S CM5 chip (GE HEALTHCARE®), respectively, using standard amine coupling methods. Cytokines were diluted to 1 and 10 μg / ml in HBS-EP+ running buffer (GE HEALTHCARE®) and injected across flowcells 1 and 2. Buffer-only injections across flowcells 1 and 2 were included for double-referencing. Data analysis was performed using BIACORE™ T200 evaluation software. The resulting sensorgrams were double-referenced (test flowcell values subtracted from control flowcell values and also a buffer blank).

[0202] Antibody 3A5.046 was found to bind only to human IL-5 and not human cytokines IL-2, IL-3, IL-4, GM-CSF, M-CSF, or GH (FIG. 11A-FIG. 11D), thus demonstrating high specificity of 3A5.046 for human IL-5.Evaluation of Antibody 3A5.046 in an Ascaris suum-Induced Cynomolgus Monkey Airways Eosinophilia Model

[0203] Antibody 3A5.046 was tested using the Ascaris suum cynomolgus model of acute airways eosinophilia as described in Hart, T. K., et al., Preclinical efficacy and safety of mepolizumab (SB-240563), a humanized monoclonal antibody to IL-5, in cynomolgus monkeys. J Allergy Clin Immunol, 2001. 108(2): p. 250-7.

[0204] Sixteen female cynomolgus macaques that had been previously sensitized to Ascaris suum were assessed for level of retained sensitivity by intradermal challenge with A. suum. Fourteen of these animals were selected for participation in the study based on the skin wheal response to challenge and randomized into two groups of n=7 per group. Two weeks after intradermal skin wheal testing, baseline blood and bronchoalveolar lavage fluid (BALF) samples were collected (Day 8). About 24 hours after baseline sample collection, animals were treated with either placebo (comprising aqueous buffer pH 6.3 with 200 mM arginine and 25 mM histidine) or antibody 3A5.046 (10 mg / kg intravenously, in aqueous buffer pH 6.3 with 200 mM arginine and 25 mM histidine) in a blinded manner (Day 7). One week after treatment with placebo or antibody a second intradermal A. suum challenge was performed and an A. suum inhalation challenge was given to all animals by delivery of an equal amount of A. suum extract (Greer Laboratories Inc.; 5 mg / ml in sterile water) administered via PARI® LC nebulizer (Day 0). Skin wheal diameter was measured 15 minutes after challenge. Two days afterA. suum inhalation challenge, blood and BALF samples were collected. Additional blood samples for haematology endpoints and PK analysis were collected weekly starting on Day 10 until Day 45.

[0205] There was no significant difference in the skin wheal responses between the first and second intradermal A. suum challenges. All animals showed an increase in eosinophils in the BALF as a result of A. suum challenge but in animals treated with antibody 3A5.046, this increase was negligible. There were significantly fewer eosinophils in the BALF of animals treated with antibody 3A5.046, compared to placebo-treated animals 48 hours after A. suum challenge (p<0.01; Mann-Whitney test; FIG. 12).

[0206] A. suum challenge also led to significantly increased blood eosinophils over baseline in placebo-treated animals to day 10 post-challenge (FIG. 13A). Treatment with antibody 3A5.046 led to a significant decrease in blood eosinophils to levels below baseline (FIG. 13A), and this depression in blood eosinophil numbers was maintained over a period of at least 52 days after initial dose (FIG. 13B).

[0207] In summary, treatment with 3A5.046 significantly reduced A. suum-induced airways eosinophilia 48 hours after challenge, and significantly decreased blood eosinophils, to below baseline levels, for at least 52 days following a single intravenous (IV) 10 mg / kg dose.Investigation of Antibody Function in a Humanized IL-5 Rat Model of Airways Eosinophilia

[0208] Alternaria alternata (Alternaria) is a fungal allergen that is known to trigger asthma in humans and can cause eosinophilia and asthma-like symptoms in rodents. When challenged with Alternaria, humanized IL-5 knock-in rats (animals in which the rat IL-5 has been replaced by human IL-5) show elevated eosinophil levels in bronchoalveolar lavage fluid (BALF) from lungs (FIG. 14).

[0209] A single intratracheal instillation of Alternaria suspension is administered to humanized IL-5 rats, and blood and BALF samples are collected 2, 3 or 4 days after Alternaria administration. Typically in this model eosinophils are significantly increased in BALF samples and may be either increased or unchanged in blood samples. Test antibodies are administered prior to induction of eosinophilia with Altemaria and assessed for their ability to reduce eosinophil numbers in BALF and blood.

[0210] Those skilled in the art will appreciate that numerous changes and modifications can be made to the preferred embodiments of the invention and that such changes and modifications can be made without departing from the spirit of the invention. It is, therefore, intended that the appended claims cover all such equivalent variations as fall within the true spirit and scope of the invention.

[0211] The disclosures of each patent, patent application, and publication cited or described in this document are hereby incorporated herein by reference, in its entirety.

[0212] TABLE 15SequencesProtein chainSequenceProtein ChainSequenceConsensus LCDR1GX1X2X3X4X5X6KX7X8YSEQ ID NO: 1Consensus LCDR2DDX8X9RPSSEQ ID NO: 2Consensus LCDR3QVWX10SSSDX11VX12SEQ ID NO: 3Antibody 3A53A5 HCDR1GGSISNGGYYWS3A5 LCDR1GGNNIGSKNVYSEQ ID NO: 4SEQ ID NO: 53A5 HCDR2YIYYSGSTY3A5 LCDR2DDSDRPSSEQ ID NO: 6SEQ ID NO: 73A5 HCDR3LGNWFDY3A5 LCDR3QVWYSSSDHVVSEQ ID NO: 8SEQ ID NO: 93A5 VHQVQLQESGPGLVKPSQTLSLTCTVSGGSISNGGYYWSWIRQHPGKGLEWIGYIYYSGSTYYNPSLKSRVSISVDTSKNQFSLKLNSVTAADTAVYYCASLGNWFDYWGQGTLVTVSSSEQ ID NO: 103A5 VLSSILTQPPSVSVAPGQTARITCGGNNIGSKNVYWYQQKPGQAPVLVVHDDSDRPSGIPERFSGSNSGNTATLTISRVEVGDEADYSCQVWYSSSDHVVFGGGTKLTVLGSEQ ID NO: 113A5 HCQVQLQESGPGLVKPSQTLSLTCTVSGGSISNGGYYWSWIRQHPGKGLEWIGYIYYSGSTYYNPSLKSRVSISVDTSKNQFSLKLNSVTAADTAVYYCASLGNWFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGSEQ ID NO: 123A5 LCSSILTQPPSVSVAPGQTARITCGGNNIGSKNVYWYQQKPGQAPVLVVHDDSDRPSGIPERFSGSNSGNTATLTISRVEVGDEADYSCQVWYSSSDHVVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECSSEQ ID NO: 13Antibody 3A5.0013A5.001 HCDR1GGSISNGGYYWS3A5.001 LCDR1GGNNIGSKNVYSEQ ID NO: 4SEQ ID NO: 53A5.001 HCDR2YIYYSGSTY3A5.001 LCDR2DDSDRPSSEQ ID NO: 6SEQ ID NO: 73A5.001 HCDR3LGNWFDY3A5.001 LCDR3QVWYSSSDHVVSEQ ID NO: 8SEQ ID NO: 93A5.001 VHQVQLQESGPGLVKPSQTLSLTCTVSGGSISNGGYYWSWIRQHPGKGLEWIGYIYYSGSTYYNPSLKSRVSISVDTSKNQFSLKLNSVTAADTAVYYCASLGNWFDYWGQGTLVTVSSSEQ ID NO: 103A5.001 VLSSILTQPPSVSVAPGQTARITCGGNNIGSKNVYWYQQKPGQAPVLVVHDDSDRPSGIPERFSGSNSGNTATLTISRVEVGDEADYSCQVWYSSSDHVVFGGGTKLTVLGSEQ ID NO: 113A5.001 HCQVQLQESGPGLVKPSQTLSLTCTVSGGSISNGGYYWSWIRQHPGKGLEWIGYIYYSGSTYYNPSLKSRVSISVDTSKNQFSLKLNSVTAADTAVYYCASLGNWFDYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGSEQ ID NO: 143A5.001 LCSSILTQPPSVSVAPGQTARITCGGNNIGSKNVYWYQQKPGQAPVLVVHDDSDRPSGIPERFSGSNSGNTATLTISRVEVGDEADYSCQVWYSSSDHVVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECSSEQ ID NO: 13Antibody 3A5.0403A5.040 HCDR1GGSISNGGYYWS3A5.040 LCDR1GGNNIGSKNVYSEQ ID NO: 4SEQ ID NO: 53A5.040 HCDR2YIYYSGSTY3A5.040 LCDR2DDSDRPSSEQ ID NO: 6SEQ ID NO: 73A5.040 HCDR3LGNWFDY3A5.040 LCDR3QVWDSSSDHVVSEQ ID NO: 8SEQ ID NO: 153A5.040 VHQVQLQESGPGLVKPSQTLSLTCTVSGGSISNGGYYWSWIRQHPGKGLEWIGYIYYSGSTYYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCASLGNWFDYWGQGTLVTVSSSEQ ID NO: 163A5.040 VLSYVLTQPPSVSVAPGQTARITCGGNNIGSKNVYWYQQKPGQAPVLVVHDDSDRPSGIPERFSGSNSGNTATLTISRVEVGDEADYSCQVWDSSSDHVVFGGGTKLTVLGSEQ ID NO: 173A5.040 HCQVQLQESGPGLVKPSQTLSLTCTVSGGSISNGGYYWSWIRQHPGKGLEWIGYIYYSGSTYYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCASLGNWFDYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGSEQ ID NO: 183A5.040 LCSYVLTQPPSVSVAPGQTARITCGGNNIGSKNVYWYQQKPGQAPVLVVHDDSDRPSGIPERFSGSNSGNTATLTISRVEVGDEADYSCQVWDSSSDHVVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECSSEQ ID NO: 19Antibody 3A5.0463A5.046 HCDR1GGSISNGGYYWS3A5.046 LCDR1GGNNIGSKNVYSEQ ID NO: 4SEQ ID NO: 53A5.046 HCDR2YIYYSGSTY3A5.046 LCDR2DDSDRPSSEQ ID NO: 6SEQ ID NO: 73A5.046 HCDR3LGNWFDY3A5.046 LCDR3QVWDSSSDHVVSEQ ID NO: 8SEQ ID NO: 153A5.046 VHQVQLQESGPGLVKPSQTLSLTCTVSGGSISNGGYYWSWIRQHPGKGLEWIGYIYYSGSTYYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCASLGNWFDYWGQGTLVTVSSSEQ ID NO: 163A5.046 VLSYVLTQPPSVSVAPGQTARITCGGNNIGSKNVYWYQQKPGQAPVLVVHDDSDRPSGIPERFSGSNSGNTATLTISRVEVGDEADYSCQVWDSSSDHVVFGGGTKLTVLGSEQ ID NO: 173A5.046 HCQVQLQESGPGLVKPSQTLSLTCTVSGGSISNGGYYWSWIRQHPGKGLEWIGYIYYSGSTYYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCASLGNWFDYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGSEQ ID NO: 203A5.046 LCSYVLTQPPSVSVAPGQTARITCGGNNIGSKNVYWYQQKPGQAPVLVVHDDSDRPSGIPERFSGSNSGNTATLTISRVEVGDEADYSCQVWDSSSDHVVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECSSEQ ID NO: 19Antibody 3A5.0633A5.063 HCDR1GGSISNGGYYWS3A5.063 LCDR1GKNNIGSKNVYSEQ ID NO: 4SEQ ID NO: 213A5.063 HCDR2YIYYSGSTY3A5.063 LCDR2DDSDRPSSEQ ID NO: 6SEQ ID NO: 73A5.063 HCDR3LGNWFDY3A5.063 LCDR3QVWDSSSDHVVSEQ ID NO: 8SEQ ID NO: 153A5.063 VHQVQLQESGPGLVKPSQTLSLTCTVSGGSISNGGYYWSWIRQHPGKGLEWIGYIYYSGSTYYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCASLGNWFDYWGQGTLVTVSSSEQ ID NO: 163A5.063 VLSYVLTQPPSVSVAPGQTARITCGKNNIGSKNVYWYQQKPGQAPVLVVHDG25KDSDRPSGIPERFSGSNSGNTATLTISRVEVGDEADYSCQVWDSSSDHVVFGGGTKLTVLGSEQ ID NO: 223A5.063 HCQVQLQESGPGLVKPSQTLSLTCTVSGGSISNGGYYWSWIRQHPGKGLEWIGYIYYSGSTYYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCASLGNWFDYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGSEQ ID NO: 183A5.063 LCSYVLTQPPSVSVAPGQTARITCGKNNIGSKNVYWYQQKPGQAPVLVVHDG25KDSDRPSGIPERFSGSNSGNTATLTISRVEVGDEADYSCQVWDSSSDHVVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECSSEQ ID NO: 23Antibody 3A5.0703A5.070 HCDR1GGSISNGGYYWS3A5.070 LCDR1GGDNIGSKNVYSEQ ID NO: 4SEQ ID NO: 243A5.070 HCDR2YIYYSGSTY3A5.070 LCDR2DDSDRPSSEQ ID NO: 6SEQ ID NO: 73A5.070 HCDR3LGNWFDY3A5.070 LCDR3QVWDSSSDHVVSEQ ID NO: 8SEQ ID NO: 153A5.070 VHQVQLQESGPGLVKPSQTLSLTCTVSGGSISNGGYYWSWIRQHPGKGLEWIGYIYYSGSTYYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCASLGNWFDYWGQGTLVTVSSSEQ ID NO: 163A5.070 VLSYVLTQPPSVSVAPGQTARITCGGDNIGSKNVYWYQQKPGQAPVLVVHDN26DDSDRPSGIPERFSGSNSGNTATLTISRVEVGDEADYSCQVWDSSSDHVVFGGGTKLTVLGSEQ ID NO: 253A5.070 HCQVQLQESGPGLVKPSQTLSLTCTVSGGSISNGGYYWSWIRQHPGKGLEWIGYIYYSGSTYYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCASLGNWFDYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGSEQ ID NO: 183A5.070 LCSYVLTQPPSVSVAPGQTARITCGGDNIGSKNVYWYQQKPGQAPVLVVHDN26DDSDRPSGIPERFSGSNSGNTATLTISRVEVGDEADYSCQVWDSSSDHVVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECSSEQ ID NO: 26Antibody 3A5.0823A5.082 HCDR1GGSISNGGYYWS3A5.082 LCDR1GGNHIGSKNVYSEQ ID NO: 4SEQ ID NO: 273A5.082 HCDR2YIYYSGSTY3A5.082 LCDR2DDSDRPSSEQ ID NO: 6SEQ ID NO: 73A5.082 HCDR3LGNWFDY3A5.082 LCDR3QVWDSSSDHVVSEQ ID NO: 8SEQ ID NO: 153A5.082 VHQVQLQESGPGLVKPSQTLSLTCTVSGGSISNGGYYWSWIRQHPGKGLEWIGYIYYSGSTYYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCASLGNWFDYWGQGTLVTVSSSEQ ID NO: 163A5.082 VLSYVLTQPPSVSVAPGQTARITCGGNHIGSKNVYWYQQKPGQAPVLVVHDN27HDSDRPSGIPERFSGSNSGNTATLTISRVEVGDEADYSCQVWDSSSDHVVFGGGTKLTVLGSEQ ID NO: 283A5.082 HCQVQLQESGPGLVKPSQTLSLTCTVSGGSISNGGYYWSWIRQHPGKGLEWIGYIYYSGSTYYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCASLGNWFDYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGSEQ ID NO: 183A5.082 LCSYVLTQPPSVSVAPGQTARITCGGNHIGSKNVYWYQQKPGQAPVLVVHDN27HDSDRPSGIPERFSGSNSGNTATLTISRVEVGDEADYSCQVWDSSSDHVVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECSSEQ ID NO: 29Antibody 3A5.0843A5.084 HCDR1GGSISNGGYYWS3A5.084 LCDR1GGNNAGSKNVYSEQ ID NO: 4SEQ ID NO: 303A5.084 HCDR2YIYYSGSTY3A5.084 LCDR2DDSDRPSSEQ ID NO: 6SEQ ID NO: 73A5.084 HCDR3LGNWFDY3A5.084 LCDR3QVWDSSSDHVVSEQ ID NO: 8SEQ ID NO: 153A5.084 VHQVQLQESGPGLVKPSQTLSLTCTVSGGSISNGGYYWSWIRQHPGKGLEWIGYIYYSGSTYYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCASLGNWFDYWGQGTLVTVSSSEQ ID NO: 163A5.084 VLSYVLTQPPSVSVAPGQTARITCGGNNAGSKNVYWYQQKPGQAPVLVVHI28ADDSDRPSGIPERFSGSNSGNTATLTISRVEVGDEADYSCQVWDSSSDHVVFGGGTKLTVLGSEQ ID NO: 313A5.084 HCQVQLQESGPGLVKPSQTLSLTCTVSGGSISNGGYYWSWIRQHPGKGLEWIGYIYYSGSTYYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCASLGNWFDYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGSEQ ID NO: 183A5.084 LCSYVLTQPPSVSVAPGQTARITCGGNNAGSKNVYWYQQKPGQAPVLVVHI28ADDSDRPSGIPERFSGSNSGNTATLTISRVEVGDEADYSCQVWDSSSDHVVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECSSEQ ID NO: 32Antibody 3A5.1073A5.107 HCDR1GGSISNGGYYWS3A5.107 LCDR1GGNNIGKKNVYSEQ ID NO: 4SEQ ID NO: 333A5.107 HCDR2YIYYSGSTY3A5.107 LCDR2DDSDRPSSEQ ID NO: 6SEQ ID NO: 73A5.107 HCDR3LGNWFDY3A5.107 LCDR3QVWDSSSDHVVSEQ ID NO: 8SEQ ID NO: 153A5.107 VHQVQLQESGPGLVKPSQTLSLTCTVSGGSISNGGYYWSWIRQHPGKGLEWIGYIYYSGSTYYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCASLGNWFDYWGQGTLVTVSSSEQ ID NO: 163A5.107 VLSYVLTQPPSVSVAPGQTARITCGGNNIGKKNVYWYQQKPGQAPVLVVHDS30KDSDRPSGIPERFSGSNSGNTATLTISRVEVGDEADYSCQVWDSSSDHVVFGGGTKLTVLGSEQ ID NO: 343A5.107 HCQVQLQESGPGLVKPSQTLSLTCTVSGGSISNGGYYWSWIRQHPGKGLEWIGYIYYSGSTYYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCASLGNWFDYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGSEQ ID NO: 183A5.107 LCSYVLTQPPSVSVAPGQTARITCGGNNIGKKNVYWYQQKPGQAPVLVVHDS30KDSDRPSGIPERFSGSNSGNTATLTISRVEVGDEADYSCQVWDSSSDHVVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECSSEQ ID NO: 35Antibody 3A5.1253A5.125 HCDR1GGSISNGGYYWS3A5.125 LCDR1GGNNIGSKHVYSEQ ID NO: 4SEQ ID NO: 363A5.125 HCDR2YIYYSGSTY3A5.125 LCDR2DDSDRPSSEQ ID NO: 6SEQ ID NO: 73A5.125 HCDR3LGNWFDY3A5.125 LCDR3QVWDSSSDHVVSEQ ID NO: 8SEQ ID NO: 153A5.125 VHQVQLQESGPGLVKPSQTLSLTCTVSGGSISNGGYYWSWIRQHPGKGLEWIGYIYYSGSTYYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCASLGNWFDYWGQGTLVTVSSSEQ ID NO: 163A5.125 VLSYVLTQPPSVSVAPGQTARITCGGNNIGSKHVYWYQQKPGQAPVLVVHDN32HDSDRPSGIPERFSGSNSGNTATLTISRVEVGDEADYSCQVWDSSSDHVVFGGGTKLTVLGSEQ ID NO: 373A5.125 HCQVQLQESGPGLVKPSQTLSLTCTVSGGSISNGGYYWSWIRQHPGKGLEWIGYIYYSGSTYYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCASLGNWFDYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGSEQ ID NO: 183A5.125 LCSYVLTQPPSVSVAPGQTARITCGGNNIGSKHVYWYQQKPGQAPVLVVHDN32HDSDRPSGIPERFSGSNSGNTATLTISRVEVGDEADYSCQVWDSSSDHVVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECSSEQ ID NO: 38Antibody 3A5.1273A5.127 HCDR1GGSISNGGYYWS3A5.127 LCDR1GGNNIGSKNAYSEQ ID NO: 4SEQ ID NO: 393A5.127 HCDR2YIYYSGSTY3A5.127 LCDR2DDSDRPSSEQ ID NO: 6SEQ ID NO: 73A5.127 HCDR3LGNWFDY3A5.127 LCDR3QVWDSSSDHVVSEQ ID NO: 8SEQ ID NO: 153A5.127 VHQVQLQESGPGLVKPSQTLSLTCTVSGGSISNGGYYWSWIRQHPGKGLEWIGYIYYSGSTYYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCASLGNWFDYWGQGTLVTVSSSEQ ID NO: 163A5.127 VLSYVLTQPPSVSVAPGQTARITCGGNNIGSKNAYWYQQKPGQAPVLVVHDV33ADSDRPSGIPERFSGSNSGNTATLTISRVEVGDEADYSCQVWDSSSDHVVFGGGTKLTVLGSEQ ID NO: 403A5.127 HCQVQLQESGPGLVKPSQTLSLTCTVSGGSISNGGYYWSWIRQHPGKGLEWIGYIYYSGSTYYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCASLGNWFDYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGSEQ ID NO: 183A5.127 LCSYVLTQPPSVSVAPGQTARITCGGNNIGSKNAYWYQQKPGQAPVLVVHDV33ADSDRPSGIPERFSGSNSGNTATLTISRVEVGDEADYSCQVWDSSSDHVVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECSSEQ ID NO: 41Antibody 3A5.1613A5.161 HCDR1GGSISNGGYYWS3A5.161 LCDR1GGNNIGSKNVYSEQ ID NO: 4SEQ ID NO: 53A5.161 HCDR2YIYYSGSTY3A5.161 LCDR2DDLDRPSSEQ ID NO: 6SEQ ID NO: 423A5.161 HCDR3LGNWFDY3A5.161 LCDR3QVWDSSSDHVVSEQ ID NO: 8SEQ ID NO: 153A5.161 VHQVQLQESGPGLVKPSQTLSLTCTVSGGSISNGGYYWSWIRQHPGKGLEWIGYIYYSGSTYYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCASLGNWFDYWGQGTLVTVSSSEQ ID NO: 163A5.161 VLSYVLTQPPSVSVAPGQTARITCGGNNIGSKNVYWYQQKPGQAPVLVVHDS52LDLDRPSGIPERFSGSNSGNTATLTISRVEVGDEADYSCQVWDSSSDHVVFGGGTKLTVLGSEQ ID NO: 433A5.161 HCQVQLQESGPGLVKPSQTLSLTCTVSGGSISNGGYYWSWIRQHPGKGLEWIGYIYYSGSTYYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCASLGNWFDYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGSEQ ID NO: 183A5.161 LCSYVLTQPPSVSVAPGQTARITCGGNNIGSKNVYWYQQKPGQAPVLVVHDS52LDLDRPSGIPERFSGSNSGNTATLTISRVEVGDEADYSCQVWDSSSDHVVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECSSEQ ID NO: 44Antibody 3A5.1693A5.169 HCDR1GGSISNGGYYWS3A5.169 LCDR1GGNNIGSKNVYSEQ ID NO: 4SEQ ID NO: 53A5.169 HCDR2YIYYSGSTY3A5.169 LCDR2DDSSRPSSEQ ID NO: 6SEQ ID NO: 453A5.169 HCDR3LGNWFDY3A5.169 LCDR3QVWDSSSDHVVSEQ ID NO: 8SEQ ID NO: 153A5.169 VHQVQLQESGPGLVKPSQTLSLTCTVSGGSISNGGYYWSWIRQHPGKGLEWIGYIYYSGSTYYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCASLGNWFDYWGQGTLVTVSSSEQ ID NO: 163A5.169 VLSYVLTQPPSVSVAPGQTARITCGGNNIGSKNVYWYQQKPGQAPVLVVHDD53SDSSRPSGIPERFSGSNSGNTATLTISRVEVGDEADYSCQVWDSSSDHVVFGGGTKLTVLGSEQ ID NO: 463A5.169 HCQVQLQESGPGLVKPSQTLSLTCTVSGGSISNGGYYWSWIRQHPGKGLEWIGYIYYSGSTYYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCASLGNWFDYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGSEQ ID NO: 183A5.169 LCSYVLTQPPSVSVAPGQTARITCGGNNIGSKNVYWYQQKPGQAPVLVVHDD53SDSSRPSGIPERFSGSNSGNTATLTISRVEVGDEADYSCQVWDSSSDHVVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECSSEQ ID NO: 47Antibody 3A5.2323A5.232 HCDR1GGSISNGGYYWS3A5.232 LCDR1GGNNIGSKNVYSEQ ID NO: 4SEQ ID NO: 53A5.232 HCDR2YIYYSGSTY3A5.232 LCDR2DDSDRPSSEQ ID NO: 6SEQ ID NO: 73A5.232 HCDR3LGNWFDY3A5.232 LCDR3QVWLSSSDHVVSEQ ID NO: 8SEQ ID NO: 483A5.232 VHQVQLQESGPGLVKPSQTLSLTCTVSGGSISNGGYYWSWIRQHPGKGLEWIGYIYYSGSTYYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCASLGNWFDYWGQGTLVTVSSSEQ ID NO: 163A5.232 VLSYVLTQPPSVSVAPGQTARITCGGNNIGSKNVYWYQQKPGQAPVLVVHDD92LDSDRPSGIPERFSGSNSGNTATLTISRVEVGDEADYSCQVWLSSSDHVVFGGGTKLTVLGSEQ ID NO: 493A5.232 HCQVQLQESGPGLVKPSQTLSLTCTVSGGSISNGGYYWSWIRQHPGKGLEWIGYIYYSGSTYYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCASLGNWFDYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGSEQ ID NO: 183A5.232 LCSYVLTQPPSVSVAPGQTARITCGGNNIGSKNVYWYQQKPGQAPVLVVHDD92LDSDRPSGIPERFSGSNSGNTATLTISRVEVGDEADYSCQVWLSSSDHVVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECSSEQ ID NO: 50Antibody 3A5.2763A5.276 HCDR1GGSISNGGYYWS3A5.276 LCDR1GGNNIGSKNVYSEQ ID NO: 4SEQ ID NO: 53A5.276 HCDR2YIYYSGSTY3A5.276 LCDR2DDSDRPSSEQ ID NO: 6SEQ ID NO: 73A5.276 HCDR3LGNWFDY3A5.276 LCDR3QVWDSSSDSVVSEQ ID NO: 8SEQ ID NO: 513A5.276 VHQVQLQESGPGLVKPSQTLSLTCTVSGGSISNGGYYWSWIRQHPGKGLEWIGYIYYSGSTYYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCASLGNWFDYWGQGTLVTVSSSEQ ID NO: 163A5.276 VLSYVLTQPPSVSVAPGQTARITCGGNNIGSKNVYWYQQKPGQAPVLVVHDH95bSDSDRPSGIPERFSGSNSGNTATLTISRVEVGDEADYSCQVWDSSSDSVVFGGGTKLTVLGSEQ ID NO: 523A5.276 HCQVQLQESGPGLVKPSQTLSLTCTVSGGSISNGGYYWSWIRQHPGKGLEWIGYIYYSGSTYYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCASLGNWFDYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGSEQ ID NO: 183A5.276 LCSYVLTQPPSVSVAPGQTARITCGGNNIGSKNVYWYQQKPGQAPVLVVHDH95bSDSDRPSGIPERFSGSNSGNTATLTISRVEVGDEADYSCQVWDSSSDSVVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECSSEQ ID NO: 53Antibody 3A5.2783A5.278 HCDR1GGSISNGGYYWS3A5.278 LCDR1GGNNIGSKNVYSEQ ID NO: 4SEQ ID NO: 53A5.278 HCDR2YIYYSGSTY3A5.278 LCDR2DDSDRPSSEQ ID NO: 6SEQ ID NO: 73A5.278 HCDR3LGNWFDY3A5.278 LCDR3QVWDSSSDYVVSEQ ID NO: 8SEQ ID NO: 543A5.278 VHQVQLQESGPGLVKPSQTLSLTCTVSGGSISNGGYYWSWIRQHPGKGLEWIGYIYYSGSTYYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCASLGNWFDYWGQGTLVTVSSSEQ ID NO: 163A5.278 VLSYVLTQPPSVSVAPGQTARITCGGNNIGSKNVYWYQQKPGQAPVLVVHDH95bYDSDRPSGIPERFSGSNSGNTATLTISRVEVGDEADYSCQVWDSSSDYVVFGGGTKLTVLGSEQ ID NO: 553A5.278 HCQVQLQESGPGLVKPSQTLSLTCTVSGGSISNGGYYWSWIRQHPGKGLEWIGYIYYSGSTYYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCASLGNWFDYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGSEQ ID NO: 183A5.278 LCSYVLTQPPSVSVAPGQTARITCGGNNIGSKNVYWYQQKPGQAPVLVVHDH95bYDSDRPSGIPERFSGSNSGNTATLTISRVEVGDEADYSCQVWDSSSDYVVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECSSEQ ID NO: 56Antibody 3A5.2793A5.279 HCDR1GGSISNGGYYWS3A5.279 LCDR1GGNNIGSKNVYSEQ ID NO: 4SEQ ID NO: 53A5.279 HCDR2YIYYSGSTY3A5.279 LCDR2DDSDRPSSEQ ID NO: 6SEQ ID NO: 73A5.279 HCDR3LGNWFDY3A5.279 LCDR3QVWDSSSDDVVSEQ ID NO: 8SEQ ID NO: 573A5.279 VHQVQLQESGPGLVKPSQTLSLTCTVSGGSISNGGYYWSWIRQHPGKGLEWIGYIYYSGSTYYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCASLGNWFDYWGQGTLVTVSSSEQ ID NO: 163A5.279 VLSYVLTQPPSVSVAPGQTARITCGGNNIGSKNVYWYQQKPGQAPVLVVHDH95bDDSDRPSGIPERFSGSNSGNTATLTISRVEVGDEADYSCQVWDSSSDDVVFGGGTKLTVLGSEQ ID NO: 583A5.279 HCQVQLQESGPGLVKPSQTLSLTCTVSGGSISNGGYYWSWIRQHPGKGLEWIGYIYYSGSTYYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCASLGNWFDYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGSEQ ID NO: 183A5.279 LCSYVLTQPP SVSVAPGQTARITCGGNNIGSKNVYWYQQKPGQAPVLVVHDH95bDDSDRPSGIPERFSGSNSGNTATLTISRVEVGDEADYSCQVWDSSSDDVVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECSSEQ ID NO: 59Antibody 3A5.2943A5.294 HCDR1GGSISNGGYYWS3A5.294 LCDR1GGNNIGSKNVYSEQ ID NO: 4SEQ ID NO: 53A5.294 HCDR2YIYYSGSTY3A5.294 LCDR2DDSDRPSSEQ ID NO: 6SEQ ID NO: 73A5.294 HCDR3LGNWFDY3A5.294 LCDR3QVWDSSSDHVASEQ ID NO: 8SEQ ID NO: 603A5.294 VHQVQLQESGPGLVKPSQTLSLTCTVSGGSISNGGYYWSWIRQHPGKGLEWIGYIYYSGSTYYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCASLGNWFDYWGQGTLVTVSSSEQ ID NO: 163A5.294 VLSYVLTQPP SVSVAPGQTARITCGGNNIGSKNVYWYQQKPGQAPVLVVHDV97ADSDRPSGIPERFSGSNSGNTATLTISRVEVGDEADYSCQVWDSSSDHVAFGGGTKLTVLGSEQ ID NO: 613A5.294 HCQVQLQESGPGLVKPSQTLSLTCTVSGGSISNGGYYWSWIRQHPGKGLEWIGYIYYSGSTYYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCASLGNWFDYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGSEQ ID NO: 183A5.294 LCSYVLTQPPSVSVAPGQTARITCGGNNIGSKNVYWYQQKPGQAPVLVVHDV97ADSDRPSGIPERFSGSNSGNTATLTISRVEVGDEADYSCQVWDSSSDHVAFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECSSEQ ID NO: 62Antibody 3A5.3023A5.302 HCDR1GGSISNGGYYWS3A5.302 LCDR1GGNNIGSKNVYSEQ ID NO: 4SEQ ID NO: 53A5.302 HCDR2YIYYSGSTY3A5.302 LCDR2DDSDRPSSEQ ID NO: 6SEQ ID NO: 73A5.302 HCDR3LGNWFDY3A5.302 LCDR3QVWDSSSDHVWSEQ ID NO: 8SEQ ID NO: 633A5.302 VHQVQLQESGPGLVKPSQTLSLTCTVSGGSISNGGYYWSWIRQHPGKGLEWIGYIYYSGSTYYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCASLGNWFDYWGQGTLVTVSSSEQ ID NO: 163A5.302 VLSYVLTQPPSVSVAPGQTARITCGGNNIGSKNVYWYQQKPGQAPVLVVHDV97WDSDRPSGIPERFSGSNSGNTATLTISRVEVGDEADYSCQVWDSSSDHVWFGGGTKLTVLGSEQ ID NO: 643A5.302 HCQVQLQESGPGLVKPSQTLSLTCTVSGGSISNGGYYWSWIRQHPGKGLEWIGYIYYSGSTYYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCASLGNWFDYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGSEQ ID NO: 183A5.302 LCSYVLTQPPSVSVAPGQTARITCGGNNIGSKNVYWYQQKPGQAPVLVVHDV97WDSDRPSGIPERFSGSNSGNTATLTISRVEVGDEADYSCQVWDSSSDHVWFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECSSEQ ID NO: 653A5.0973A5.097 HCDR1GGSISNGGYYWS3A5.097 LCDR1GGNNIDSKNVYSEQ ID NO: 4SEQ ID NO: 663A5.097 HCDR2YIYYSGSTY3A5.097 LCDR2DDSDRPSSEQ ID NO: 6SEQ ID NO: 73A5.097 HCDR3LGNWFDY3A5.097 LCDR3QVWDSSSDHVVSEQ ID NO: 8SEQ ID NO: 153A5.097 VHQVQLQESGPGLVKPSQTLSLTCTVSGGSISNGGYYWSWIRQHPGKGLEWIGYIYYSGSTYYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCASLGNWFDYWGQGTLVTVSSSEQ ID NO: 163A5.097 VLSYVLTQPPSVSVAPGQTARITCGGNNIDSKNVYWYQQKPGQAPVLVVHDG29DDSDRPSGIPERFSGSNSGNTATLTISRVEVGDEADYSCQVWDSSSDHVVFGGGTKLTVLGSEQ ID NO: 673A5.097 HCQVQLQESGPGLVKPSQTLSLTCTVSGGSISNGGYYWSWIRQHPGKGLEWIGYIYYSGSTYYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCASLGNWFDYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGSEQ ID NO: 183A5.097 LCSYVLTQPPSVSVAPGQTARITCGGNNIDSKNVYWYQQKPGQAPVLVVHDG29DDSDRPSGIPERFSGSNSGNTATLTISRVEVGDEADYSCQVWDSSSDHVVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECSSEQ ID NO: 683A5.046 VHCAGGTGCAGCTGCAGGAATCTGGCCCTGGCCTGGTCAAGCCCAGCCAnucleotide GACCCTGAGCCTGACCTGTACCGTGTCCGGCGGCAGCATCAGCAACGGsequenceCGGCTACTACTGGTCCTGGATCAGACAGCACCCCGGCAAGGGCCTGG(synthetic)AATGGATCGGCTACATCTACTACAGCGGCAGCACCTACTACAACCCCAGCCTGAAGTCCAGAGTGACCATCAGCGTGGACACCAGCAAGAACCAGTTCAGCCTGAAGCTGAGCAGCGTGACAGCCGCCGACACCGCCGTGTACTACTGCGCCAGCCTGGGCAATTGGTTCGACTACTGGGGCCAGGGCACCCTCGTGACAGTGTCCTCASEQ ID NO: 693A5.046 human HCGCTAGCACCAAGGGCCCCAGCGTGTTCCCCCTGGCCCCTTGTAGCAGAconstantAGCACCAGCGAGAGCACAGCCGCCCTGGGCTGCCTGGTGAAAGACTAregionCTTCCCCGAGCCCGTCACCGTGTCCTGGAACAGCGGAGCCCTGACCAGnucleotideCGGCGTGCACACCTTTCCAGCCGTGCTGCAGAGCAGCGGCCTGTACAGsequenceCCTGAGCAGCGTGGTGACAGTGCCCTCCAGCAGCCTGGGCACCAAGA(synthetic)CCTACACCTGTAACGTGGACCACAAGCCCAGCAACACCAAGGTGGACAAGCGGGTGGAATCTAAGTACGGCCCACCCTGCCCCCCCTGCCCTGCCCCTGAATTTCTGGGCGGACCCTCCGTGTTCCTGTTCCCCCCAAAGCCCAAGGACACCCTGTATATCACTCGGGAGCCCGAAGTGACCTGCGTGGTGGTGGACGTGTCCCAGGAAGATCCCGAGGTCCAGTTCAATTGGTACGTGGACGGCGTGGAAGTGCACAACGCCAAGACCAAGCCCAGAGAGGAACAGTTCAACAGCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGACTGGCTGAACGGCAAAGAGTACAAGTGCAAAGTCTCCAACAAGGGCCTGCCCAGCTCCATCGAGAAAACCATCAGCAAGGCCAAGGGCCAGCCCCGCGAGCCTCAGGTGTACACACTGCCCCCCAGCCAGGAAGAGATGACCAAGAACCAGGTGTCCCTGACCTGTCTGGTGAAAGGCTTCTACCCCAGCGATATCGCCGTGGAATGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCTGTGCTGGACAGCGACGGCAGCTTCTTCCTGTACTCCCGGCTGACCGTGGACAAGAGCCGGTGGCAGGAAGGCAACGTCTTCAGCTGCAGCGTGATGCACGAGGCCCTGCACAACCACTACACCCAGAAGTCCCTGAGCCTGAGCCTGGGCSEQ ID NO: 703A5.046 VLAGCTACGTGCTGACCCAGCCTCCTAGCGTGTCCGTGGCCCCTGGCCAGnucleotideACCGCCAGAATCACCTGTGGCGGCAACAACATCGGCAGCAAGAACGTsequenceGTACTGGTATCAGCAGAAGCCCGGCCAGGCCCCCGTGCTGGTGGTGCA(synthetic)CGACGACAGCGACAGACCCAGCGGCATCCCCGAGCGGTTCAGCGGCAGCAACAGCGGCAATACCGCCACCCTGACCATCAGCCGGGTGGAAGTGGGCGACGAGGCCGACTACAGCTGCCAGGTCTGGGACAGCAGCAGCGACCACGTGGTGTTCGGCGGAGGCACCAAGCTGACCGTCCTAGGTSEQ ID NO: 713A5.046 human LCCAGCCCAAGGCCGCTCCCAGCGTGACCCTGTTCCCCCCAAGCAGCGAGconstantGAACTGCAGGCCAACAAGGCCACCCTGGTGTGCCTGATCAGCGACTTCregionTACCCTGGGGCCGTGACCGTGGCCTGGAAGGCCGATAGCAGCCCTGTGnucleotideAAGGCCGGCGTGGAAACCACCACCCCCTCCAAGCAGAGCAACAACAAsequenceATACGCCGCCAGCAGCTACCTGTCCCTGACCCCCGAGCAGTGGAAGTC(synthetic)CCACCGGTCCTACAGCTGCCAGGTGACACACGAGGGCAGCACCGTGGAAAAGACCGTGGCCCCCACCGAGTGCAGCSEQ ID NO: 72EMBODIMENTS

[0213] The following list of embodiments is intended to complement, rather than displace or supersede, the previous descriptions.

[0214] Embodiment 1. A human antibody molecule that immunospecifically binds to human IL-5 with an equilibrium affinity constant (KD) of at least about 40 pM as determined by surface plasmon resonance.

[0215] Embodiment 2. A human antibody molecule that immunospecifically binds to human IL-5, the antibody molecule comprising:

[0216] a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO: 6, a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO: 8, a light chain CDR1 comprising the amino acid sequence of SEQ ID NOs: 5, 21, 24, 27, 30, 33, 36, 39, or 66, a light chain CDR2 comprising the amino acid sequence of SEQ ID NOs: 7, 42, or 45, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NOs: 15, 48, 51, 54, 57, 60, or 63.

[0217] Embodiment 3. The antibody molecule of embodiment 1 or 2, wherein the antibody molecule comprises a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO: 6, a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO: 8, and

[0218] a. a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 5, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 15;

[0219] b. a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 21, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 15;

[0220] c. a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 24, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 15;

[0221] d. a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 27, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 15;

[0222] e. a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 30, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 15;

[0223] f. a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 33, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 15;

[0224] g. a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 36, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 15;

[0225] h. a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 39, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 15;

[0226] i. a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 66, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 15;

[0227] j. a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 5, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 42, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 15;

[0228] k. a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 5, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 45, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 15;

[0229] l. a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 5, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 48;

[0230] m. a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 5, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 51;

[0231] n. a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 5, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 54;

[0232] o. a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 5, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 57;

[0233] p. a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 5, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 60; or

[0234] q. a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 5, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 63;

[0235] wherein the position of the amino acid residues of the CDR is determined according to AbM.

[0236] Embodiment 4. The antibody molecule of embodiment 3, wherein the antibody molecule comprises a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO: 6, a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO: 8, a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 5, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 15.

[0237] Embodiment 5. The antibody molecule of any one of the previous embodiments, wherein the antibody molecule comprises a heavy chain variable region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 16 and

[0238] a. a light chain variable region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 17;

[0239] b. a light chain variable region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 22;

[0240] c. a light chain variable region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 25;

[0241] d. a light chain variable region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 28;

[0242] e. a light chain variable region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 31;

[0243] f. a light chain variable region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 34;

[0244] g. a light chain variable region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 37;

[0245] h. a light chain variable region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 40;

[0246] i. a light chain variable region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 43;

[0247] j. a light chain variable region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 46;

[0248] k. a light chain variable region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 49;

[0249] l. a light chain variable region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 52;

[0250] m. a light chain variable region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 55;

[0251] n. a light chain variable region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 58;

[0252] o. a light chain variable region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 61;

[0253] p. a light chain variable region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 64; or

[0254] q. a light chain variable region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 67.

[0255] Embodiment 6. The antibody molecule of any one of the previous embodiments, wherein the antibody molecule comprises a heavy chain variable region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 16 and a light chain variable region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 17.

[0256] Embodiment 7. The antibody molecule of any one of the previous embodiments, wherein the antibody molecule comprises:

[0257] a. a S228P mutation;

[0258] b. a M252Y mutation, a S254T mutation, and a T256E mutation;

[0259] c. a deletion of a heavy chain C-terminal lysine residue; or

[0260] d. any combination of a to c.

[0261] Embodiment 8. The antibody molecule of embodiment 7, wherein the antibody molecule comprises a S228P mutation and a deletion of a heavy chain C-terminal lysine residue.

[0262] Embodiment 9. The antibody molecule of embodiment 7, wherein the antibody molecule comprises a S228P mutation, a M252Y mutation, a S254T mutation, a T256E mutation, and a deletion of a heavy chain C-terminal lysine residue.

[0263] Embodiment 10. The antibody molecule of any one of the previous embodiments, wherein the antibody molecule comprises an IgG4 heavy chain constant region and a lambda light chain constant region.

[0264] Embodiment 11. The antibody molecule of any one of the previous embodiments, wherein the antibody molecule comprises a heavy chain comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 18 and

[0265] a. a light chain comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 19;

[0266] b. a light chain comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 23;

[0267] c. a light chain comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 26;

[0268] d. a light chain comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 29;

[0269] e. a light chain comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 32;

[0270] f. a light chain comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 35;

[0271] g. a light chain comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 38;

[0272] h. a light chain comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 41;

[0273] i. a light chain comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 44;

[0274] j. a light chain comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 47;

[0275] k. a light chain comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 50;

[0276] l. a light chain comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 53;

[0277] m. a light chain comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 56;

[0278] n. a light chain comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 59;

[0279] o. a light chain comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 62;

[0280] p. a light chain comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 65; or

[0281] q. a light chain comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 68.

[0282] Embodiment 12. The antibody molecule of any one of embodiments 1-10, wherein the antibody molecule comprises a heavy chain comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to

[0283] the amino acid sequence of SEQ ID NO: 20 and a light chain comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 19.

[0284] Embodiment 13. The antibody molecule of embodiment 2 or embodiment 3, wherein the antibody molecule is a Fab fragment, a Fab2 fragment, or a single chain antibody.

[0285] Embodiment 14. The antibody molecule of any one of the previous embodiments, wherein the antibody molecule has one or more of the following properties:

[0286] a. reduces binding of IL-5 to the IL-5 receptor;

[0287] b. has a serum half-life of at least about 20 days; or

[0288] c. binds human and cynomolgus monkey IL-5 but not mouse, rat, or guinea pig IL-5.

[0289] Embodiment 15. The antibody molecule of embodiment 2, wherein the antibody molecule binds to human IL-5 with an equilibrium affinity constant (KD) of at least about 40 pM as determined by surface plasmon resonance.

[0290] Embodiment 16. A pharmaceutical composition comprising the antibody molecule of any one of embodiments 1 to 15.

[0291] Embodiment 17. A nucleic acid molecule encoding the antibody molecule of any one of embodiments 1 to 15.

[0292] Embodiment 18. A vector comprising the nucleic acid molecule of embodiment 17.

[0293] Embodiment 19. A cell transformed to express the antibody molecule of any one of embodiments 1 to 15.

[0294] Embodiment 20. A method of treating a subject having eosinophilic asthma, hypereosinophilic syndrome, nasal polyposis with eosinophilic involvement, eosinophilic granulomatosis with polyangiitis, atopic dermatitis, or eosinophilic esophagitis comprising:

[0295] administering to the subject a therapeutically effective amount of the antibody molecule of any one of embodiments 1 to 15 or the pharmaceutical composition of embodiment 16 to treat the eosinophilic asthma, hypereosinophilic syndrome, nasal polyposis with eosinophilic involvement, eosinophilic granulomatosis with polyangiitis, atopic dermatitis or eosinophilic esophagitis.

[0296] Embodiment 21. Use of an effective amount of the antibody molecule of any one of embodiments 1 to 15 or the pharmaceutical composition of embodiment 16 in the treatment of eosinophilic asthma, hypereosinophilic syndrome, nasal polyposis with eosinophilic involvement, eosinophilic granulomatosis with polyangiitis, atopic dermatitis, or eosinophilic esophagitis.

[0297] Embodiment 22. Use of the antibody molecule of any one of embodiments 1 to 15 or the pharmaceutical composition of embodiment 16 in the manufacture of a medicament for the treatment of eosinophilic asthma, hypereosinophilic syndrome, nasal polyposis with eosinophilic involvement, eosinophilic granulomatosis with polyangiitis, atopic dermatitis or eosinophilic esophagitis.

Examples

examples

[0153]The following examples are provided to further describe some of the embodiments disclosed herein. The examples are intended to illustrate, not to limit, the disclosed embodiments.

Generation of Anti-Human-IL-5 Antibodies

[0154]Anti-human-IL-5 antibodies were obtained from transgenic rats (OMT) with human V-genes cloned into their genomes and which produce antibodies with human V-domains and rat Fc domains. Briefly, the transgenic rats were genetically immunized with DNA encoding IL-5 four times over 21 days (on days 0, 7, 14, 21) and boosted with recombinant human IL-5 at day 28 of the immunization protocol. Serum antibody titres were determined at days 0 and 38 of the immunization protocol by an ELISA assay using recombinant human IL-5. Briefly, sera from each animal were diluted in PBS 1% BSA and were tested using ELISA plates coated with 1 μg / ml human IL-5, or BSA as a control. A goat anti-rat IgG R-phycoerythrin conjugate (SOUTHERN BIOTECH®, #3030-09) was used at 10 μg / ml as...

embodiments

[0213]The following list of embodiments is intended to complement, rather than displace or supersede, the previous descriptions.[0214]Embodiment 1. A human antibody molecule that immunospecifically binds to human IL-5 with an equilibrium affinity constant (KD) of at least about 40 pM as determined by surface plasmon resonance.[0215]Embodiment 2. A human antibody molecule that immunospecifically binds to human IL-5, the antibody molecule comprising:[0216]a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO: 6, a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO: 8, a light chain CDR1 comprising the amino acid sequence of SEQ ID NOs: 5, 21, 24, 27, 30, 33, 36, 39, or 66, a light chain CDR2 comprising the amino acid sequence of SEQ ID NOs: 7, 42, or 45, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NOs: 15, 48, 51, 54, 57, 60, or 63.[0217]Embodiment 3. The antibody ...

Claims

1. A human antibody molecule that immunospecifically binds to human Interleukin-5 (IL-5), wherein the antibody molecule comprises:a heavy chain complementarity determining region 1 (CDR1) comprising the amino acid sequence of SEQ ID NO: 4, a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO: 6, a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO: 8, a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 5, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 15, andan IgG4 heavy chain comprising a S228P mutation, a M252Y mutation, a S254T mutation, a T256E mutation, and a deletion of a heavy chain C-terminal lysine residue.

2. The antibody molecule of claim 1, wherein the antibody molecule comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 16 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 17.

3. The antibody molecule of claim 1, wherein the antibody molecule comprises the IgG4 heavy chain constant region comprising the S228P mutation, the M252Y mutation, the S254T mutation, the T256E mutation, and the deletion of the heavy chain C-terminal lysine residue, and a lambda light chain constant region.

4. The antibody molecule of claim 3, wherein the antibody molecule comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 20 and a light chain comprising the amino acid sequence of SEQ ID NO: 19.

5. A pharmaceutical composition comprising the antibody molecule of claim 1.

6. A pharmaceutical composition comprising the antibody molecule of claim 4.

7. A nucleic acid molecule encoding the antibody molecule of claim 1.

8. A vector comprising the nucleic acid molecule of claim 7.

9. An isolated cell transformed to express the antibody molecule of claim 1.

10. A nucleic acid molecule encoding the antibody molecule of claim 4.

11. A vector comprising the nucleic acid molecule of claim 10.

12. An isolated cell transformed to express the antibody molecule of claim 4.

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